KENDRIYA VIDYALAYA SANGATHAN, NEW DELHI

ZONAL INSTITUTE OF EDUCATION AND TRAINING, GWALIOR

INTERACTIVE STUDY MATERIAL FOR CLASS XII

(BIOLOGY)

Smt Shajila P, Training Associate (Biology)

1
 INDEX
Sl. No. Content Page No.

1 Unit-wise Weightage 3

2 Reproduction in Organisms 4

3 Sexual Reproduction in Flowering Plants 8

4 Human Reproduction 17

5 Reproductive Health 26

6 Principles of Inheritance 29

7 Molecular Basis of Inheritance 40

8 Evolution 55

9 Human Health and Diseases 61

10 Microbes in Human Welfare 76

11 Biotechnology- Principles and Processes 79

12 Biotechnology and its Applications 88

13 Organisms and Populations 96

14 Ecosystem 101

15 Biodiversity and Conservation 109

2
 CLASS XII- BIOLOGY (2022-23)

Unit-Wise Weightage:

Unit Title Marks

Reproduction
VI Chapter 2: Sexual Reproduction in Flowering Plants 14
Chapter-3: Human Reproduction
Chapter 4: Reproductive Health

Genetics an Evolution
VII Chapter-5: Principles of Inheritance and Variation 18
Chapter-6: Molecular Basis of Inheritance
Chapter-7: Evolution

Biology and Human Welfare

VIII Chapter-8: Human Health and Disease 14
Chapter-10: Microbes in Human Welfare

Biotechnology and its Applications

IX Chapter-11: Biotechnology: Principles and Processes 12
Chapter-12: Biotechnology and its Applications

Ecology and Environment

X Chapter-13: Organisms and Populations 12
Chapter-14: Ecosystem
Chapter-15: Biodiversity and its Conservation

Total 70

Please click the link:

For Class XII Syllabus for 2022-23:
https://drive.google.com/file/d/1URNn1b7o1XPYRi12BnilDzZmBurKSqJt/view?usp=share_link
For sample paper:
https://drive.google.com/file/d/1L0mpnsZ51Dvs2nSbtbBH14HNsSvEOHXM/view?usp=sharing
https://drive.google.com/file/d/1E-XcKQBEXqWPEltVy201VqF_QPQkLHGz/view?usp=sharing
3
 UNIT VI - REPRODUCTION
CHAPTER 1
REPRODUCTION IN ORGANISMS
Note: This chapter is deleted for the year 2022-23. Here it is given as an introduction for the unit.
Reproduction:
Defined as a biological process in which an organism gives rise to young ones (offspring)
similar to itself.

Phases of Life (Life Span of Organisms)

Juvenile Phase Reproductive Phase Senescence phase

PPT link for further explanations

https://docs.google.com/presentation/d/1Y_pLa7PkWt6fnI9tg0_1dOz7iWamXjpa/edit?usp=share_link&ouid=107171444
976724466844&rtpof=true&sd=true

Reproduction

Asexual Reproduction Sexual Reproduction

Fission - eg. Amoeba Eye buds in tuber

Budding - eg. Hydra Rhizome of ginger

Conidia – eg. Penicillium Bulbil of Agave

Zoospore - eg. Leaf buds of Bryophyllum

Gemmule – eg. Sponge Offset of water hyacinth

Vegetative Propagation Bulb in onion

4
 Asexual reproduction Sexual reproduction
Offsprings are produced by single parents Offsprings are produced by the involvement of
single or two parents
Does not involve gamete formation and Gamete formation and gamete fusion takes place.
gamete fusion
No zygote formation Gametes are fused to form a diploid zygote (2n)
and zygote develops into embryo.

Offsprings are genetically identical to their Offsprings are not genetically identical to their
parents parents

Asexual reproduction
Methods of asexual reproduction:
 Binary fission; e.g. Amoeba, Paramoecium.
 Budding: e.g. yeast.
 Asexual reproductive structures:
Zoospores: aquatic fungi, eg. Chlamydomonas.
Conidia: Penicillium.
Bud:Hydra
Gemmules: sponges.
 Vegetative propagation units in plant: (Vegetative propagules)
Rrhizome, tuber, offset, bulb.etc.

For more examples -

https://drive.google.com/file/d/17zJdXsZwRx9M9faQlHee9Ar3cbkNZ_WH/view?usp=sharing

5
Sexual reproduction
Features of sexual reproduction:

Events in Sexual Reproduction

Pre fertilisation Fertilisation Post fertilisation

Gametogenesis Internal Zygote

Gamete transfer External Embryogenesis

Pre-fertilization events:

Gametogenesis: Process of gamete formation.

There are two types of gametes- isogametes: (when two gametes are morphologically similar) and
heterogametes: (when the two gametes produced are morphologically different)
Male gamete is called antherozoids or sperm and the female gamete is called ovum or egg.
There are two types of plants based on their sexuality.
i) Monoecious Plants: Possess both male and female sex organs.
ii) Dioecious Plants: Possess only one sex organ, male or female.

Gamete transfer: During this event both male and female gametes are physically brought
together to facilitate fertilization.

* In most cases male gametes are motile, female gametes are non-motile.
* In case of few fungi and algae, both male and female gametes are motile.
* In most cases water is the medium for gamete transfer.
* Male gametes are produced in several thousand times the number of female gametes produced to
compensate the loss during transfer.

6
Types of Gametes

Isogametes of Cladophora

Heterogametes of Fucus

Heterogametes of Homo sapiens

Fertilization:
* Fusion of male and female gamete is called fertilization or syngamy.
* Two types of fertilization- Internal (Gametic fusion takes place inside the body) and External
(Gametic fusion takes place outside the body)
* The female gamete undergoes development to form new organism without fertilization. This
phenomenon is called parthenogenesis.

Post Fertilisation Events:

Zygote formation: Fusion of male and female gametes results in the formation of zygote.

Embryogenesis: Development of zygote into an embryo is called embryogenesis.

Zygote undergoes cell division (mitosis) and cell differentiation. Cell division increase the number of
cells and cell differentiation helps to form specialized tissues and organs.
*****************************

7
 CHAPTER 2
SEXUAL REPRODUCTION IN FLOWERING PLANTS

Structure of Flower

Calyx (Consists of Sepals) Androecium Gynoecium Corolla (Consists of Petals)

Stamen Filament Stigma Style Ovary

Pre fertilization events:

This stage is characterised by the production of male and female gametes and transfer of
gametes for fertilization process. During this stage androecium (stamen) and gynoecium (pistil)
are fully developed for the production of gametes.

Structure of Stamen:
Typical stamen consists of two parts, long and slender stalk called filament and terminal bilobed structure
called anther.

8
Anther:
Atypical angiosperm anther is a bilobed and dithecous structure. Each anther contains four
microsporangia located at the corners, two in each lobe. Microsporangia become pollen sacs and are
packed with pollen grains.

Structure of microsporangium:
Each microsporangium surrounded by four wall layers
* Epidermis
* Endothecium Protects the inner tissues.
* Middle layer.
* Tapetum. Nourishes the developing pollen grain.
The center of each microsporangium contains homogenous cells called sporogenous tissues.

Microsporogenesis:
* The process of formation of microspores from pollen mother cell through meiosis is called
microsporogenesis.
* The sporogenous tissue of microsporangium differentiated into microspore mother cell or pollen
mother cell.
* Each microspore mother cell undergoes meiosis and gives rise to haploid microspore tetrad.
* On dehydration microspore tetrad dissociated to form four microspores.
* Each microspore developed into a pollen grain.

Video link for Microsporogenesis

https://drive.google.com/file/d/1oARpDAhFiVjnCFHWyBcKcsIHf4lEvNyr/view?usp=share_link

https://www.youtube.com/watch?v=Vus-lgBrIV8

9
 PPT link for Microsporogenesis and structure of pollen grain
https://docs.google.com/presentation/d/1m7C99e6twj35bFdRTjHwm1d6eXzjd98O/edit?usp=sharing&ouid=107
171444976724466844&rtpof=true&sd=true

(a
(b

(a) Stages of a microspore maturing into a pollen grain (b) Enlarged view of pollen grain tetrad

The Pistil:
Pistil has three parts the stigma, style and ovary. Inside the ovary is the ovarian cavity (locule).
Megasporongia or ovules arise from the placenta. The number of ovule inside the ovary may be single or
many.

The Megasporangium (Ovule):

Megasporangium is attached to the placenta of locule with a stalk called funicle. The body of the ovule
fused with the funicle in the region called hilum. Hilum is the junction between the funicle and ovule.
Each ovule has one or two protective envelops called integuments with an opening called micropyle.
Opposite of the micropylar end, is the chalaza, representing the basal part of the ovule.

10
 Pistil Anatropous Ovule

Megasporogenesis:
* The process of formation of megaspores from the megaspore mothercell is called
Megasporogenesis.
* In the centre of the ovule there is a mass of tissue called nucellus.
* Cells of nucellus have abundant reserve food materials.
* One cell of the nucellus towards micropylar end differentiated into megaspore mother cell
(MMC).
* It is a large diploid cell, dense cytoplasm with prominent nucleus.
* The MMC undergo meiotic division resulting four haploid megaspores.

PPT Link for Megasporogenesis

https://docs.google.com/presentation/d/10Cpgs8y17bGxC-
BQKv89sws2BQROAQQ9/edit?usp=sharing&ouid=107171444976724466844&rtpof=true&sd=true

11
Pollination:
Transfer of pollen grains from the anther to the stigma of a pistil is termed as pollination.

Kinds of pollination:

Autogamy Geitonogamy Xenogamy

Pollination within same Pollination between two Transfer of pollen grains from the anther
flower. flowers of the same plant. to the stigma of different plant.

Self-Pollination Pollination by pollinating It is commonly called as cross-

agent. pollination.

Genetically similar plants are Genetically similar to the It brings genetically different types of
produced autogamy. pollen grains to the stigma and
genetically different plants are produced.

Viola, Oxalis and Commelina produce two types of flowers:

Chasmogamous: With exposed anther and stigma
Cleistogamous: With closed anther and stigma.
Cleistogamous flower is invariably autogamous and assured seed set even in the absence of the
pollinator.

Agents of pollination:

Anemophily Hydrophily Entomophily

Pollinating agent is wind. Pollination by water. Pollinating agent is insect
particularly bees are the
dominant biotic agents for
pollination.

Produces more number of pollen Vallisneria,Hydrilla and flowers are very large,
to compensate the uncertainties Zostera are the common colorful, fragrant and rich in
of pollination. example for Hydrophily. nectar.

Flowers with well exposed Pollen grains released into Small flowers present in
stamens. the surface of water and cluster to make them
carried to the stigma by air conspicuous.
current as in Vallisneria.

Large feathery stigma to trap air- This type of pollination is Flower pollinated by flies and
borne pollen grains. very rare, mostly monocot. beetles secrete foul odours.
In sea grass the flowers
remained submerged.

Contains single ovule in one Pollen grains are long, Contains single ovule in one
ovary and numerous flower ribbon like and carried ovary and

12
 packed into an inflorescence passively inside the water
e.g. corn cob.

Pollen grains are light and non- Pollen grains are protected
sticky. from wetting by
mucilaginous covering.

Outbreeding Devices:
* Pollen released and stigma receptivity is not synchronized.
* Spatial separation of anthers and stigmas
* Anther and stigma are placed at different positions.
* Self incompatibility.
* Production of unisexual flowers.

PPT Link for Double fertilisation

https://drive.google.com/file/d/1KHyulpTc-cWR8hfwy4Un7R4JRR6PNG8Z/view?usp=sharing

Video link for Double fertilization

https://www.youtube.com/embed/dgFY7WUTASQ

13
 Pollen pistil Interaction: All the events – from pollen deposition on the stigma until pollen tubes
enter the ovule – are together referred as pollen-pistil interaction.

The pistil has the ability to recognize the pollen

whether it is compatible or incompatible.

If it is right type the stigma allow the pollen to

germinate.

The pollen grain produce pollen tube through one of

the germ pore.

Pollen tube grows through the tissues of the stigma

and style and reaches the ovary.

Contents of the pollen grain move into the pollen

tube.

The generative cell divides and forms two male

gametes inside the pollen tube.

Pollen tube enters into the ovule through micropyle

and then into the embryo sac through synergids guided
by filiform apparatus.

Double fertilization:
* After entering one of the synergids, the pollen tube releases two male gametes into the cytoplasm of
the synergids.
* Syngamy: one of the male gamete fused with egg cell, to form a diploid zygote.
* Two polar nuclei of central cell fused to form a diploid secondary nucleus.

14
 * Triple fusion: The second male gamete fused with the secondary nucleus to form a triploid
primary endosperm nucleus.
* Since two type of fusion, syngamy and triple fusion take place in the embryo sac the phenomenon is
termed as double fertilization.
* The central cell after triple fusion becomes primary endosperm cell and developed into the
endosperm.
* The zygote developed into an embryo.

Longitudinal section of a pistil showing growth of pollen tube

POST- FERTILIZATION : STRUCTURE AND EVENTS

Events of endosperm and embryo development, maturation of ovule into seed and ovary into fruit, are
collectively termed as post-fertilization events.

Endosperm:
* Development of endosperm takes place before the embryo development.
* Primary endosperm cell divides repeatedly to form a triploid endosperm.
* PEN undergoes successive nuclear division to give rise to free nuclei. This is called free-nuclear
endosperm.
Embryo:
* Zygote formed and placed at the micropylar end of the embryo sac and undergoes cell division and
develop into embryo.

15
APOMIXIS AND POLYEMBRYONY.
 Apomixis is very common in Asteraceae and grasses.
 Seeds are produced without fertilization.
 Apomixis is a type of asexual reproduction which mimics the sexual reproduction.
 Diploid egg cell is formed without meiosis and develops into seed without fertilization.
 In Citrusand Mango the nucellar cells starts dividing, protrude into the embryo sac and
develop into embryo.
 Ovule having more than one embryo is termed as polyembryony.
 Hybrid plants are developed by apomixis to maintain the genetic identity.

********************************************************************

16
 CHAPTER 3
HUMAN REPRODUCTION
THE MALE REPRODUCTIVE SYSTEM.

Male reproductive system includes a pair of testes, accessory ducts, accessory glands and external
genitalia

Testes:
Testes is located outside the abdominal cavity within a pouch called scrotum. Scrotum provides low
temperature required for spermatogenesis. Each testis has about 250 compartments called testicular
lobules. Each lobule contains one to three seminiferous tubules. Male germ cell undergoes meiosis
and produce sperm. Sertoli cells provide nutrition to the germ cell and the sperm. In between the
seminiferous tubule there is interstitial cell or Leydig cells that produce testicular hormones called
androgen (testosterone).

17
 Hormones involved in the regulation of Spermatogenesis

THE FEMALE REPRODUCTIVE SYSTEM

The female reproductive system includes a pair of ovaries, a pair of oviduct, uterus and
cervix, vagina, external genitalia and a pair of mammary gland.

18
Oviduct:
* Oviducts, uterus and vagina constitute the female accessory ducts.
* Each fallopian tube is about 10-12 cm long and extends from the periphery of each ovary to the
uterus.
* Close to the ovary the oviduct has a funnel shaped structure called infundibulum?
* The edges of the infundibulum possess finger-like projections called fimbriae, which helps in
collection of the ovum after ovulation.
* The infundibulum leads to a wider part of the oviduct called ampulla.
* The last part of the oviduct is called isthmus which joined to uterus.
Uterus:
* Attached the pelvic wall by ligaments.
* The uterus opens into vagina through a narrow cervix.
* The lumen of cervix is called cervical canal.
* Cervical canal along with vagina form the birth canal.
* The wall of the uterus has three layers of tissues Perimetrium, Myometrium and Endometrium.
* Endometrium undergoes cyclical changes during menstrual cycle.
* Myometrium exhibits strong contraction during delivery of the baby.

GAMETOGENESIS: (formation of gametes)

Spermatogenesis:
Link for Spermatogenesis:
https://drive.google.com/file/d/1fiTw5LSOkXlRu_FbodaTJIAYRrwKAs9x/view?usp=sharing

Sectional view of seminiferous tubule

19
 Hormonal control of spermatogenesis:
* This process is initiated at puberty due to secretion of gonadotrophins releasing hormone (GnRH)
* GnRH secreted form hypothalamus and stimulate anterior pituitary to secrete two gonadotrophins.
Luteinizing hormone (LH) and Follicle stimulating Hormone (FSH).
* LH acts on Leydig cells and stimulates synthesis of androgens.
* Androgen stimulates spermatogenesis.
* FSH acts on Sertoli cells and stimulates spermatogenesis in other ways.

Structure of sperm:
* Ultrastructure of sperm consists of a head, neck, a middle piece and a tail.
* Whole body of sperm surrounded by plasma membrane.
* The sperm head contain an elongated haploid nucleus.
* Above the nucleus a cap like structure present called acrosome.
* The acrosome contains enzymes which help in fertilization of ovum.
* The middle piece contains mitochondria, which provide energy for movement of tail that facilitate
sperm motility.
* Sperm released from seminiferous tubules enters into accessory ducts.
* On their way fluids from seminal vesicle and prostate gland added which collectively called as
Semen.

Link for spermatogenesis- Video: https://www.youtube.com/embed/6fBa8UqEano

Structure of Sperm
Oogenesis:
Formation of a mature female gamete or ovum is called oogenesis.
Link for Oogenesis:
https://drive.google.com/file/d/14nIrhCzc8ZFALyjVcBU7YWvqggbTrSRc/view?usp=sharing
Link for oogenesis- Video: https://www.youtube.com/embed/hKa57JPfKDE

20
 Gametogenesis

Menstrual cycle:
* Reproductive cycle of female primates is called menstrual cycle.
* The first menstruation begins at puberty is called Menarche.
* Menstrual cycle repeated at an average interval of 28/29 days.
* One ovum is released in the middle of each menstrual cycle.

21
Video of Menstrual cycle
https://www.youtube.com/embed/ZNM8KsSQerk

Menstrual cycle has four phases:

i) Menstrual phase, ii) Follicular phase, iii)Ovulatory phase and iv)Luteal phase.

Fertlisation:

Fertilisation

Sex determination:
* Sex of a baby has been decided during fertilization and in the zygote.
* Sex is determined by the sex-chromosomes present in gametes.
* Human female contain two XX chromosomes.
* Human male contain XY chromosomes.
* All the female gametes produced with only ‘X’ chromosome.
* Sperms produced by male, 50% with ‘X’ and 50 % with ‘Y’ chromosome.
* After fertilization zygote either carries XX or XY chromosomes.
* Zygote with XX chromosomes develop into female and with XY chromosome develops into male.

Sex determination in Human beings

22
Cleavage: Repeated mitotic division of the zygote without growth resulting a multicellular ball like
embryo is called cleavage.

Zygote

Cleavage

Morula

Blastula (Blastocyst)

Trophoblast Inner cell mass

Helps development of Embryo

placenta

Cleavage and formation of blastocyst

23
 Implantation: The attachment of blastocyst in the uterine endometrium is called Implantation..

Pregnancy and embryonic development:

* After implantation, finger like projections appears on the trophoblast called chorionic villi.
* Chorionic villi surrounded by uterine tissue and maternal blood.
* chorionic villi and uterine tissue become interdigitated with each other and jointly form a structural
and functional unitbetween developing embryo (foetus) and maternal body called placenta.
Function of placenta:
* The embryo connected to the placenta by umbilical cord, which transports substances to and from
the embryo.
* Facilitate transport of oxygen and nutrient from mother to embryo.
* Removes CO2 and waste material from the embryo.
* Acts as endocrine gland and produces several hormones like: Human chorionic gonadotrophins
(hCG), Human placental lactogen (hPL), Estrogen, Progesterone and Relaxin produced from the
ovary in the later stage of pregnancy.

Embryonic development:
* After implantation the inner cell mass of blastocyst differentiated into an outer layer called
ectoderm and an inner layer called endoderm.
* Mesoderm differentiated in-between ectoderm and endoderm.
* The inner cell mass thus called stem cells, having potency to produce all types of cell, tissues and
organs by differentiation.
Organogenesis:
* Formation of different organs in the embryo is called organogenesis.
* Human pregnancy lasts for 9 months.
* After one month of pregnancy heart is formed in the embryo.
* By the end of 2nd month the foetus develops limbs and digits.
* By the end of 12 weeks (first trimester) most of organ system is formed (limbs and external
genitalia are well developed).
* First movement of foetus and appearance of hairs observed in 5th month.
* By the end of 24th week (2nd trimesters) the body is covered with fine hairs, eye-lids separate,
and eyelashes are formed.
* By the end of 9 months the foetus is fully developed and is ready for delivery.

24
PARTURATION:

The period of pregnancy is called gestation period. (9 months).

 Ejection or expulsion or delivery of foetus is called parturition.
 Parturition is due to vigorous contraction of uterine Myometrium.
 The signal of parturition is originated from the fully developed foetus and the placenta
which induces mild contraction of uterus called fetal ejection reflex.
 Fetal ejection reflex triggers the release of Oxytocin from pituitary.
 Oxytocin induces stronger contraction of uterine endometrium.
 Stimulatory reflex continues stronger contraction leads to expulsion.
 After delivery the placenta is also expelled out of the uterus.

****************************************************************************

25
 CHAPTER 4
REPRODUCTIVE HEALTH
Abbreviations:

Abbreviations Expanded form

IUCD Intra Uterine Contraceptive Device

RCH Reproductive and Child Health care

STD Sexually Transmitted Disease

HIV Human Immuno deficiency virus.

AIDS Acquired immuno deficiency syndrome

CDRI Central Drug Research Institute

MMR Maternal Mortality Rate

IMR. Infant mortality rate

MTP Medical Termination of Pregnancy

VD Venereal Disease

RTI Reproductive Tract Infection

PID Pelvic Inflammatory Disease

ART Assisted Reproductive Technologies

IVF In Vitro Fertilisation

ZIFT Zygote Intra Fallopian Transfer

AI Artificial insemination

IUI Interna uterine insemination.

26
 ET Embryo transfer.

IUT Intra uterine transfer.

ICSI Intra Cytoplasmic Sperm Injection

POPULATION EXPLOSION AND BIRTH CONTROL:

* Increased health facilities, better living conditions are the cause of population explosion.
* Out of 6 billion world population 1 billion are Indians.
* Rapid decline in death rate, maternal mortalility rate (MMR) and infant mortality rate (IMR)
are major cause of population growth.
* Indian population growth rate is around 1.7 percent.
Characteristics of ideal contraceptive.
* User friendly.
* Easily available.
* Effective
* Nor or least side – effects.
* No way interferes with sexual drive.
BIRTH CONTROL METHODS:

Natural methods: Principle of working: Avoiding chances of ovum and sperms meeting.
i) Periodic abstinence
ii) Withdrawal or coitus interruption
iii) Lactational amenorrhea
Barrier methods: Principle of working: prevents physical meeting of sperm and ovum.
Condoms,Diaphragm, cervical caps and vaults.

Intra Uterine Devices: Eg. Non-medicated IUDs e.g. Lippes loop,Copper releasing IUDs (CuT, Cu7,
Multiload 375), Hormone releasing IUDs (Progestasert, LNG-20).

Principle of working:

27
 * Increase phagocytosis of sperm within the uterus.
* Cu ion released suppresses sperm motility and fertilizing capacity of sperm.
* Hormone releasing IUDs make the uterus unsuitable for implantation and the cervix hostile to the
sperm.
Oral contraceptives: Principle of working:
* Inhibit ovulation, Inhibit implantation.
* Alter the quality of cervical mucus to prevent/retard entry of sperms.
* Saheli- a non steroidal preparation used as oral contraceptive pills.

Surgical methods:

Vasectomy Tubectomy
Sterilization process in male. Sterilization process in female.
A small part of the vas deferens is removed or A small part of the fallopian tube is removed or
tied up. tied up.

MEDICAL TERMINATION OF PREGNANCY:

Intentional or voluntary termination of pregnancy before full term is called medical termination of
pregnancy (MTP) or induced abortion.
INFERTILITY:The couple unable to produce children inspite of unprotected sex.
In case there no corrections are possible, some special technologies used to have children called
assisted reproductive technologies (ART).
Assisted reproductive technologies:
(a) In vitro fertilization:
(b) Embryo transfer: ZIFT- Zygote intra fallopian transfer.IUT- Intra Uterine transfer (embryo
with more than 8 blastomeres).
(c) Gamete intra fallopian transfer- GIFT
(d) Intra cytoplasmic sperm injection (ICSI):
(e) Artificial insemination (AI)

***************************************************************************

28
 UNIT VII – GENETICS AND EVOLUTION

CHAPTER 5
PRINCIPLES OF INHERITANCE AND VARIATION
PRINCIPLES OF INHERITANCE AND VARIATION
* Genetics: deals with the inheritance, as well as the variation of characters from parents to offsprings.
* Inheritance: is the process by which characters are passed on from parent to progeny.
* Variation: is the degree by which progeny differ from their parents.
MENDEL’S LAWS OF INHERITANCE:
* Gregor Mendel. Conducted hybridization experiments on garden peas for seven years (1856 – 1863) and proposed laws
of inheritance.
* Mendel conducted artificial pollination/cross pollination experiments using several true-breeding pea lines.
* A true breeding line is one that, having undergone continuous self-pollination for several generations.
* Mendel selected 14 true-breeding peas’ plant varieties, as pair’s which were similar except for one character with
contrasting traits.

True breeds selected by Mendel

INHERITANCE OF ONE GENE: (Monohybrid cross)

* Mendel crossed tall and dwarf pea plants to study the inheritance of one gene.
* He collected the seeds produced as a result of this cross and grew them to generate plants of the
first hybrid generation. This generation is called filial progeny or the F1.
* Mendel observed that all the F1 progeny plants ere tall, like one of its parents; none were dwarf.
* He made similar observations for the other pairs of traits – he found that the F1 always resembled
either one of the parents, and that the trait of the other parent was not seen in them.
* Mendel then self – pollinated the tall F1 plants and to his surprise found that in the F2 generation
some of the offsprings were ‘dwarf; the character that was not seen in the F1 generation was now
expressed.
th
* The proportion of plants that were dwarf was 1/4 of the F2 plants while 3/4th of the F2 plants were
tall.

29
 * The tall and dwarf traits were identical to their parental type and did not show any blending, that is
all the offsprings were either tall or dwarf, none were of in between height.
* Similar results were obtained with the other traits that he studied: only one of the parental traits was
expressed in the F1 generation while at the F2 stage both the traits were expressed in the proportion
of 3:1.
* The contrasting traits did not show any blending at either F1 or F2 stage.
Mendel’s proposition:
* Mendel proposed that something was being stably passed down, unchanged, from parent to
offspring through the gametes, over successive generations. He called these things as ‘factors’.
* Now a day we call them as genes.
* Gene is therefore are the units of inheritance.
* Genes which codes of a pair of contrasting traits are known as alleles, i.e. they are slightly different
forms of the same gene.

Terminologies used in Genetics

Genetic Definition Example
Terms

Allele Different forms of a gene, Different alleles produce different hair colors—
which producevariations in brown,blond, red, black, etc.
a genetically inherited trait.

Genes Genes are parts of DNA and Genes contain blue‐print for each individual for
carry hereditary information her orhis specific traits.
passed from parents to children.
Dominant version (allele) of a
Dominant When a child inherits dominant brown‐hair gene
gene shows itsspecific trait
form(allele) from dad, the child will have brown
even if only one parent passed
hair.
the gene to the child.
When a child inherits recessive blue‐eye gene form
Recessive Recessive gene shows its
(allele) from both mom and dad, the child will have
specific trait whenboth
blueeyes.
parents pass the gene to the
child.
Homozygous Two of the same form of a Inheriting the same blue eye gene form
gene—one frommom and from bothparents result in a homozygous
the other from dad. gene.
Heterozygous Two different forms of a gene— Inheriting different eye color gene forms from
one from mom and the other momand dad result in a heterozygous gene.
from dad are different.

Genotype Internal heredity Blue eye and brown eye have different
information that contain genotypes—oneis coded for blue and the other
genetic code. for brown.
Phenotype Both having or not having a widow’s
Outwardly expressed traits or
peak arephenotypes.
characteristics.
30
 Some genetic traits follow Mendelian Inheritance,
A simple genetic rule
Mendelian whileother genetic traits follow different
where a gene onlycomes
Inheritance inheritance patterns or rules.
in dominant or recessive
forms.

31
Law of Dominance:
 Characters are controlled by discrete units called factors.
 Factors occur in pairs.
 In a dissimilar pair of factors one member of the pair dominates (dominant) the other
(recessive).

Law of Segregation:
 The alleles do not show any blending and that both the characters are recovered as such in
the F2 generation though one of these is not seen at the F1 stage.
 The parents contain two alleles during gamete formation; the factors or alleles of a pair
segregate or separate from each other such that a gamete receives only one of the two
factors.
 Homozygous parent produces all gametes that are similar i.e contain same type of allele.
 Heterozygous parents’ produces two kinds of gametes each having one allele with equal
proportion.

Incomplete dominance:
 When a cross between two pure breed is done for one contrasting character, the F1 hybrid
phenotype dose not resemble either of the two parents and was in between the two, called
incomplete dominance.
 Inheritance of flower color in the dog flower (snapdragon or Antirrhinum sp.)is a good
example of incomplete dominance.
32
  F2 generation phenotypic ratio is 1:2:1 in stead of 3:1 as Mendelian monohybrid cross.
 Genotypic ratio of F2 generation is 1:2:1.

For Mendel’s Laws of Inheritance click the following link:

https://www.youtube.com/embed/mD0Onu2ArGA
For Deviations from Mendelian Laws click the link given below.
https://drive.google.com/file/d/1OYeU7D48jbOkouen6Mc3yiNzXhB-wKVr/view?usp=sharing

Co – dominance:
* F1 resembled either of the two parents (complete dominance).
* F1 offspring was in-between of two parents (incomplete dominance).
* F1 generation resembles both parents (co-dominance).
* Best example of co-dominance is the ABO blood grouping in human.
* ABO blood group is controlled by the gene I.
* The plasma membrane of the RBC has sugar polymers (antigen) that protrude from its surface and
the kind of sugar is controlled by the gene-I.
A B
* The gene I has three alleles I ,I and i.
A B
* The alleles I and I produce a slightly different form of sugar while allele i doesn’t produce any
sugar.
* Each person possesses any two of the three I gene alleles.
A B
* I and I are completely dominant over i.
A B
* When I , and I present together they both express their own types of sugar; this because of co-
dominance. Hence red blood cells have both A and B type sugars.

33
Multiple Alleles:
* Example of ABO blood grouping produces a good example of multiple alleles.
* There are more than two alleles. i.e. three allele governing the same character.

A single gene product may produce more than one effect:

Eg. Starch synthesis in pea seeds is controlled by one gene. It has two alleles B and b. Starch is
synthesized effectively by BB homozygote and therefore, large starch grains are produced. The ‘bb’
homozygous has less efficiency hence produce smaller grains. After maturation of the seeds, BB seeds
are round and the bb seeds are wrinkle. Heterozygous (Bb) produce round seed and so B seems to be
dominant allele, but the starch grains produced are of intermediate size. If starch grain size is
considered as the phenotype, then from this angle the alleles show incomplete dominance.

INHERITANCE OF TWO GENES: (Dihybrid Cross)

Law of independent Assortment:

When two characters (dihybrid) are combined in a hybrid, segregation of one pair of traits is
independent of the other pair of traits.

CHROMOSOMAL THEORY OF INHERITANCE:

* Proposed by Walter Sutton and Theodore Bovery in 1902.

* The behavior of chromosomes was parallel to the behavior of genes and used chromosome
movement to explain Mendel’s laws.
* Sutton united the knowledge of chromosomal segregation with Mendelian principles and called it
the chromosomal theory of inheritance.
o Chromosome and genes are present in pairs in diploid cells.
o Homologous chromosomes separate during gamete formation (meiosis)
o Fertilization restores the chromosome number to diploid condition.
o The chromosomal theory of inheritance claims that, it is the chromosomes that segregate and assort
independently.

Linkage: Physical association of genes on a chromosome.

Recombination: The generation of non-parental gene combinations.

POLYGENIC INHERITANCE:
The inheritance one trait by three or more genes are called polygenic inheritance. In a
polygenic trait the phenotype reflects the contribution of each allele i.e. the effect of each
allele is additive.
Eg. Human skin colour.

Eg: Inheritance of Human skin colour-

Link for explanations as PPT-

https://drive.google.com/file/d/14SqO36KUMrNZ9wX6axUvEJDs0CCMzDPQ/view?usp=sharing

Link for Video:

https://www.youtube.com/embed/yMN_wbRb38M
34
 (Darkest) AABBCC X aabbcc (Fairest) (P)

AaBbCc (Intermediate) (F1)

AaBbCc X AaBbCc

F2

PLEIOTROPY:
* A single gene can exhibit multiple phenotypic expression, such gene is called pleiotropic gene.
* The mechanism of pleiotropy in most cases is the effect of a gene on metabolic pathways which
contributes towards different phenotypes.
* Phenylketonuria a disease in human is an example of pleiotropy.
* This disease is caused due to mutation in the gene that code for the enzyme phenyl alanine
hydroxylase.
* Phenotypic expression characterized by: Mental retardation, reduction in hairs and reduction in
skin pigmentation.

SEX DETERMINATION:
Sex-determination of grass hopper:
* Sex-determination in grasshopper is XX-XO type.
* All egg bears one ‘X’ chromosome along with autosomes.
* Some sperms (50%) bear’s one ‘X’ chromosome and 50% do not.
* Egg fertilized with sperm (with ‘X’ chromosome) became female (22+XX).
* Egg fertilized with sperm (without ‘X’ chromosome) became male (22 + X0)
35
Sex determination in Human beings (XX-XY type):
* Bothe male and female has same number of chromosomes.
* Female have autosomes and a pair of X chromosomes. (AA+ XX)
* Male have autosomes and one large ‘X’ chromosome and one very small ‘Y-chromosomes.
(AA+XY)
* This is called male heterogammety and female homogamety.

Sex determination in birds:

* Female birds have two different sex chromosomes designated as Z and W.
* Male birds have two similar sex chromosomes and called ZZ.
* Such type of sex determination is called female heterogammety and male homogamety.
Sex determination in Honey bee:
* Sex determination in honey bee based on the number of sets of chromosomes an individual
receives.
* An offspring formed from the fertilization of a sperm and an egg developed into either queen
(female) or worker (female).
* An unfertilized egg develops as a male (drone), by means of parthenogenesis.
* The male have half the number of chromosome than that of female.
* The female are diploid having 32 chromosomes and males are haploid i.e. having 16 numbers of
chromosomes.
* This is called haplodiploid sex determination system.
* Male produce sperms by mitosis, they don not have father and thus cannot have sons, but have
grandsons.

36
Pedigree Analysis:
 Analysis of traits in several of generations of a family is called the pedigree analysis.
 In the pedigree analysis the inheritance of a particular trait is represented in the family tree
over generations.

GENETIC DISORDERS:

Hemophilia:
In this disease a single protein that is a part of the cascade of proteins involved in the clotting of blood is
affected. Due to this in an affected individual a simple cut will result in non-stop bleeding.
* Sex linked recessive disease.
* The diseases transmitted from unaffected carrier female to some of the male progeny.
* Female becoming hemophilic is extremely rare because mother of such a female at least carrier and
the father should be hemophilic.
* Affected transmits the disease only to the son not to the daughter.
* Daughter can receive the disease from both mother and father.
Sickle cell anaemia:
* This is an autosomes linked recessive trait.
* The defect is caused due to substitution of Glutamic acid (Glu) by Valine (Val) at the sixth position
of the beta globin chain of the haemoglobin molecule.
* Substitution of amino acid takes place due to the single base substitution at the sixth codon of the
beta globin gene from GAG to GUG.

37
 * Transmitted from parents to the offspring when both the parents are carrier for the gene
(heterozygous).
* This disease is controlled by single pair of allele, HbA, and HbS.
* There are three possible genotypes (HbA HbA, HbA HbS, and HbSHbS.
* Only homozygous individuals for HbS (HbS HbS) show the diseased phenotype.
* Heterozygous (HbA HbS) individuals appear unaffected but they are carrier of the disease as there
is 50 percent probability of transmission of the mutant gene to the progeny.

Phenylketonuria:
* Autosomal recessive trait.
* Inborn error of metabolism.
* The affected individual lack one enzyme called phenyl alanine hydroxylase that converts the amino
acid phenyl alanine to tyrosine.
* In the absence of the enzyme phenyl alanine accumulated and converted into phenylpyruvic acid
and other derivatives.
* Accumulation of these results in mental retardation.
* These derivatives excreted through kidney.
Chromosomal disorders:
* Caused due to absence or excess or abnormal arrangement of one or more chromosome.
* Failure of segregation of chromatids during cell division cycle results in the gain or loss of
chromosome(s), called Aneuploidy.
* Failure of cytokinesis after telophase stage of cell division results in an increase in a whole set of
chromosome in an organism and this phenomenon is called polyploidy.
Trisomy: additional copy of a chromosome may be included in an individual (2n+1).
Monosomy: an individual may lack one of any one pair of chromosomes (2n-1)
Down syndrome:
* Caused due to presence of an additional copy of the chromosome number 21 (trisomy of 21).
* This disorder was first described by Langdon Down (1866).
o Short stature with small round head.
o Furrowed tongue
38
 o Partially opened mouth
o Palm is broad with characteristic palm crease.
o Physical, psychomotor and mental development is retarded.
Klinefelter’s syndrome:
* Caused due to the presence of an additional copy of X-chromosome resulting into a
karyotype of 47, (44+XXY).
o Overall masculine development.
o Also develop feminine character (development of breast i.e. Gynaecomastia)
o Individuals are sterile.
Turner’s syndrome:
* Caused due to the absence of one of the X- chromosomes i.e. 45 (44 + X0)
* Such females are sterile as ovaries are rudimentary.
* Lack of other secondary sexual characters.

39
 CHAPTER – 6
MOLECULAR BASIS OF INHERITANCE
THE DNA:

 DNA is a long polymer of deoxyribonucleotides.

 The length of the DNA depends on, number of nucleotide pair present in it.
 Characteristics of the organism depend on the length of the DNA.

Structure of polynucleotide chain:

* A nucleotide has three component:-
i) A nitrogen base
ii)A pentose sugar ( ribose in RNA and deoxyribose in DNA)
ii)A phosphoric acid.
* There are two types of nitrogen bases:
i) Purines ( Adenine and Guanine)
ii) Pyrimidines ( Cytosine, Uracil and Thymine)
* Adenine, Guanine and Cytosine is common in RNA and DNA.
* Uracil is present in RNA and Thymine is present in DNA in place of Uracil.
* Pentose sugar is ribose in RNA and Deoxyribose in DNA.
* A nitrogen base attached to the pentose sugar at C1 of pentose sugar by N-glycosidic linkage to
form a nucleoside.
* Phosphoric acid attached to the nucleoside by Phosphodiester linkage a corresponding nucleotide
is formed. (Ribonucleotide or deoxyribonucleotides depending on the sugar unit).

40
 * Two nucleotides are joined by 3’-5’ Phosphodiester linkage to form dinucleotide.
* More than two nucleotides joined to form polynucleotide chain.
* The backbone of the polynucleotide chain is sugar and phosphate.
* Nitrogen bases linked to the sugar (ribose or deoxyribose)
* In RNA Uracil is found in place of thymine.

Central dogma of Molecular Biolog: Flow of genetic information.

41
Packaging of DNA Helix:
Distance between two conjugative base pairs is 0.34nm, the length of the DNA in a typical mammalian
cell will be 6.6 X109 bp X 0.34 X10-9 /bp, it comes about 2.2 meters.

Packaging in prokaryotes:
* They do not have definite nucleus.
* The DNA is not scattered throughout the cell.
* DNA is held together with some proteins in a region is called ‘nucleoid’.
The DNA in nucleoid is organized in large loops held be proteins.
Packaging in Eukaryotes:
* There is a set of positively charged, basic protein called Histones.
* Histones are positively charged due to rich in basic amino acids like Lysines and arginines.
* Histones are organized to form a unit of eight molecules called histone octamer.
* Negatively charged DNA wrapped around positively charged histone octamer to form a structure
called nucleosome.
* The nucleosomes are seen as ‘beads-on-string’ structure when viewed under electron microscope.
* The chromatin is packaged to form chromatin fibers that are further coiled and condensed at
metaphase stage to form chromosome.
* Packaging at higher level required additional set of proteins called Non-histone Chromosomal
(NHC) proteins.
* In a typical nucleus some loosely coiled regions of chromatin (light stained) is called euchromatin.
* The chromatin that more densely packed and stains dark are called Heterochromatin.
* Euchromatin is transcriptionally active chromatin and heterochromatin is inactive.

42
GENETIC CODE- Salient Features:

THE SEARCH OF GENETIC MATERIAL:

Transforming principle:
Experiment:

* Given by Frederick Griffith in 1928.

* His experiment based on Streptococcus pneumoniae (caused pneumonia).
* There is change in physical form of bacteria.
* There are two colonies of bacteria:
* Smooth shiny colonies called S strain.
* Rough colonies called R strain.

43
Conclusion of experiment:
* R – Strain bacteria had some how been transformed by the heat killed S-Strain bacteria.
* Some ‘transforming principle’, transferred from heat killed S-Strain bacteria, had enabled the R-
Strain to synthesize smooth polysaccharide coat and become virulent (S Strain).
* The transformation of R-Strain to S-Strain is due to transfer of Geneticmaterial.
* However the biochemical nature of genetic material was not defined from his experiment.
* Protein of heat killed S-Strain is not the genetic material
* RNA of heat killed S-Strain is not the genetic material.
* DNA of heat killed S-Strain is the genetic material, because DNA digested with DNase mixed with
R-strain unable to transform R-Strain to S-Strain.

Biochemical characterization of transforming principle:

* Biochemical nature of transforming principle was discovered by Oswald Avery, Colin Macleod
and Maclyn McCarty. (1933-44)
* They purified biomolecules (proteins, DNA and RNA) from the heat killed S cells to see which one
could transform live R cells to S cells.
* Heat killed S-Strain + protease + Live R-Strain → transformation.
* Heat killed S-Strain + RNase + Live R-Strain → transformation.
* Heat killed S-Strain + DNase + Live R-Strain → transformation.
The Genetic Material is DNA:
* ‘DNA is the genetic material’ is proved by Alfred Hershey and Martha Chase (1952).
* They worked on the virus that infects bacteria called bacteriophage.
* During normal infection the bacteriophage first attaches the bacteria cell wall and then inserts its
genetic material into the bacterial cell.
* The viral genetic material became integral part of the bacterial genome and subsequently
manufactures more virus particle using host machinery.
* Hershey and Chase worked to discover whether it was protein or DNA from the viruses that entered
the bacteria.
Experiment : (Blenders experiment)
* They grew some viruses on a medium having radioactive phosphorus and some others on medium
having radioactive sulfur.

44
 * Viruses grown in radioactive Phosphorus have radioactive DNA but not radioactive protein
because Phosphorus present in DNA not in protein.
* Viruses grown in radioactive sulfur have radioactive protein not radioactive DNA because sulfur
present in protein but not in DNA.
* Infection: radioactive phages were allowed to attach to E.coli bacteria; the phages transfer the
genetic material to the bacteria.
* Blending: the viral coats were separated from the bacteria surface by agitating them in a blender.
* Centrifugation: The virus particles were separated from the bacteria by spinning them in a
centrifuge machine.
Observation:
* Bacteria infected with viruses that had radioactive DNA were radioactive and no radioactivity in the
supernatant.
* Bacteria infected with viruses that had radioactive protein were not radioactive, but radioactivity
found in the supernatant.
Conclusion of Experiment:
* DNA is the infecting agent that made the bacteria radioactive hence DNA is the genetic
material not the protein.

Harshey and chase Experiment: Link for the video

https://www.youtube.com/embed/i45l1RslGwM

REPLICATION: THE PROCESS:

* Watson and Crick proposed a scheme for replication of DNA.
* The Original statement that “It has not escaped our notice that the specific pairing we have
postulated immediately suggests a possible copying mechanism for the genetic material (Watson
and Crick, 1953)
* The scheme suggested that the two strands would separate and act as template for the synthesis of
new complementary strands.
* New DNA molecule must have one parental strand and one new strand.
* This scheme of replication is called Semiconservative type of replication.

Experimental Proof of semiconservative nature of replication:

* It is now proved experimentally that replication is semiconservative type.
* It was first shown in Escherichia coli and subsequently in higher organism.
* Mathew Messelson and Franklin Stahl performed the following experiment in 1958.
45
Link for Semiconservative mode of DNA replication video:
https://www.youtube.com/embed/4gdWOWjioBE

STEPS OF THE EXPERIMENTS:

* They grew E.coli in 15NH4Cl medium for many generations. (15N is heavy nitrogen not
radioactive element)
* The result was that 15N was incorporated into newly synthesized DNA and other nitrogen
containing compound as well.
* This heavy DNA molecule could be distinguished from normal DNA by centrifugation in a
cesium chloride (CsCl) density gradient.
* Then they transferred the E.coli into a medium with normal 14NH4Cl and let them
grow.(E.coli divides in 20 minutes)
* They took samples at definite time intervals as the cells multiplied, and extracted the DNA
that remained as double-stranded helices.
* Various samples were separated independently on CsCl gradients to measure the densities
of DNA.
* The DNA that was extracted from the culture one generation after the transfer from 15N to
14N medium had a hybrid or intermediate density.
* DNA extracted from the culture after another generation (after 40 min.) was composed of
equal amount of this hybrid DNA and of ‘light ‘DNA.
Experiment by Taylor and colleagues:
* Used radioactive thymidine to detect distribution of newly synthesized DNA in the chromosomes.
* They performed the experiment on Vicia faba (faba beans) in 1958.
* They proved the semiconservative nature of DNA replication in eukaryotes.

TRANSCRIPTION:

46
‘The process of copying genetic information from one strand of the DNA into RNA is termed as
transcription’.
Transcription vs. Replication:
* Principle of complementarity governs the process of transcription except Adenosine of DNA
forms base pair with the Uracil instead of thymine. During replication Adenine pairs with thymine
instead of uracil.
* During replication once started the whole DNA is duplicated, where as transcription takes place
only a segment of DNA.
* In replication both strand acts as template, where as in transcription only one strand is acts as
template to synthesize RNA.
* In replication DNA copied from a DNA, where as in transcription RNA copied from the DNA.
* Only one strand is acting as template for transcription. If both strand of DNA acts as template, they
would code for RNA molecule with different sequences and in turn if they code for proteins, the
sequence of amino acids in the protein would be different. Hence one segment of DNA would be
coding for two different proteins.
* The two RNA molecules if produced from simultaneously would be complementary to each other,
hence will form double stranded RNA. This would prevent RNA translation into protein.
Transcription unit:
47
 * A transcription unit in DNA consists of three regions:
* A promoter
* The structural gene
* A terminator.
* DNA dependent RNA polymerase catalyses the polymerization in only one direction that is
5’→3’.

Process of transcription: prokaryotes.

There is a single DNA dependent RNA polymerase that catalyses transcription or synthesis of all
three types of RNAs in prokaryotes. The process of transcription completed in three steps:
Initiation, Elongation and Termination.
There are three different types of RNA polymerases in eukaryotes.
i) RNA polymerase I transcribes rRNA (28S, 18S, and 5.8S)
ii) RNA polymerase II transcribes heterogeneous nuclear RNA (hnRNA).
iii)RNA polymerase III transcribes tRNA, 5srRNA and snRNA.
 Post transcriptional processing: (occurs inside the nucleus)
(a) Splicing:
Primary transcript (hn RNA) consists of both exons (coding) and introns (non coding) and required to be
processed before translation. Therefore the introns are removed and exons are joined in a defined order.
This process is called splicing.This is catalyzed by SnRNP, introns removed as spliceosome.
(b) Capping: An unusual nucleotide called methyl guanosine triphosphate is added to the 5’ end of
hnRNA.
(c) Tailing: Adenylate residues (200-300) are added at 3’ end of hnRNA in a template independent
manner. The processed hnRNA is now called mRNA and transported out of the nucleus for translation.

48
 Fig. Transcription in eukaryotes

tRNA-the Adaptor molecule:

* The tRNA is called sRNA (soluble RNA)
* It acts as an adapter molecule.
* tRNA has an anticodon loop that base complementary to the codon.
* It has an amino acid accepterend to which it binds with amino acid.
* Each tRNA bind with specific amino acid i.e 61 types of tRNA found.
* One specific tRNA with anticodon UAC called initiator tRNA.
* There is no tRNA for stop codons. (UAA, UGA, UAG)
* The secondary structure is like clover-leaf.
* The actual structure of tRNA is compact, looks like inverted ‘L’.

Link for Transcription, Translation - From DNA to Protein- Video

https://www.youtube.com/embed/gG7uCskUOrA

Link for PPT- From DNA to Protein-

https://docs.google.com/presentation/d/19-
d1I9DyY1VE5q0rjDVPDSR7uvWeK3n9/edit?usp=share_link&ouid=107171444976724466844&rtpof=true&sd=true

TRANSLATION:
* It refers to polymerization of amino acids to form a polypeptide.
* The number and sequence of amino acids are defined by the sequence of bases in the mRNA.
* The amino acids are joined by peptide bond.
* Amino acids are activated in the presence of ATP and linked to their specific tRNA is called
charging of tRNA or aminoacylation of tRNA.
* Ribosome is the cellular factory for protein synthesis.
* Ribosome consists of structural rRNA and 80 different proteins.
* In inactive state ribosome(70S) present in two subunits:-
* Large sub unit 50S and small sub unit 30S
.
49
Initiation:
 The process of translation or protein synthesis begins with attachment of mRNA with small
subunit of ribosome.
 The ribosome binds to the mRNA at the start codon (AUG).
 AUG is recognized by the initiator tRNA.
Elongation:
 Larger subunit attached with the initiation complex.
 Larger subunit has two site ‘A’ site and ‘P’ site.
 Initiator tRNA accommodated in ‘P’ site of large subunit, the subsequent amino-acyl-tRNA
enters into the ‘A’ site.
 The sub subsequent tRNA selected according to the codon of the mRNA.
 Codon of mRNA and anticodon of tRNA are complementary to each other.
 Formation of peptide bond between two amino acids of ‘P’ and ‘A’ site, catalyzed by
ribozyme, (23S rRNA in bacteria)
 The moves from codon to codon along the mRNA called translocation.
Termination:
 Elongation continues until a stop codon arrives at ‘P’ site.
 There is no tRNA for stop codon.
 A release factor binds to the stop codon.
 Further shifting of ribosome leads to separation of polypeptide.
 An mRNA also has some additional sequences that are not translated called untranslated
regions (UTR).

Link for Transcription, Translation - From DNA to Protein- Video

https://www.youtube.com/embed/gG7uCskUOrA

Link for PPT- From DNA to Protein-

https://docs.google.com/presentation/d/19-
d1I9DyY1VE5q0rjDVPDSR7uvWeK3n9/edit?usp=share_link&ouid=107171444976724466844&rtpof=true&sd=true

50
REGULATION OF GENE EXPRESSION:
 Regulation of gene expression in eukaryotes takes place in different level:
 Transcriptional level (formation of primary transcript)
 Processing level (regulation of splicing)
 Transport of mRNA from nucleus to the cytoplasm.
 Translational level.
 In prokaryotes control of rate of transcriptional initiation is the predominant site for control
of gene expression.
 The activity of RNA polymerase at the promoter is regulated by accessory proteins, which
affects its ability to recognize the start site.
 The regulatory proteins can acts both positively (activators) or negatively (repressor)
 The regulatory proteins interact with specific region of DNA called operator, which
regulate the accessibility of RNA polymerase to promoter.

Lac operon:
 Francois Jacob and Jacque Monod first to describe a transcriptionally regulated system of
gene expression.
 A polycistronic structural gene is regulated by common promoter and regulatory genes.
Such regulation system is common in bacteria and is called operon.
 Lac operon consists of:-
o One regulator gene ( i-gene)
o Three structural genes (z,y,a)
o Operator. (binding site of repressor protein)
o Promoter.(binding site of the RNA polymerase)

51
52
HUMAN GENOMIC PROJECT:
 Genetic make-up of an organism or an individual lies in the DNA sequences.
 Two individual differs in their DNA sequences at least in some places.
 Finding out the complete DNA sequence of human genome.
 Sequencing human genome was launched in 1990.
Goals of HGP:
 Identify all the approximately 20,000 – 25,000 genes in human DNA.
 Determine the sequence of all 3 billion chemical base pairs.
 Store this information in data bases.
 Improve tools for data analysis.
 Transfer related technologies to other sectors, such as industries.
Address the ethical, legal, and social issues (ELSI) that may arise from the project.
Methodology:
 To identify all the genes that expressed as RNA referred as Expressed Sequence Tags
(ETSs).

 Simply sequencing the whole set of genome that contained all the coding and non-coding
sequence, and later assigning different regions in the sequence with functions called
Sequence Annotation.

53
  The commonly used hosts for sequencing were bacteria and yeast and vectors were called as
BAC (bacterial artificial chromosome) and YAC (yeast artificial chromosome).
DNA FINGER PRINTING:
 DNA finger printing is a very quick way to compare the DNA sequences of any two
individuals.
 DNA fingerprinting involves identifying differences in some specific regions in DNA
called repetitive DNA, because in these sequences, a small stretch of DNA is repeated
many times.
 During centrifugation the bulk DNA forms major peak and the other small peaks are called
satellite DNA.
 Depending on base composition (A:T rich or G:C rich), length of segment, and number of
repetitive units, the satellite DNA classified into many types, such as mini –satellite and
micro – satellite.
 These sequences dose not code for any proteins.
 These sequences show high degree of polymorphism and form basis of DNA fingerprinting.
 Polymorphism in DNA sequence is the basis of genetic mapping of human genome as well
as of DNA fingerprinting.
 Polymorphism (variation at genetic level) arises due to mutations.
 If an inheritable mutation is observed in a population at high frequency it is referred as DNA
polymorphism.
The process:
 DNA fingerprinting was initially developed by Alec Jeffreys.
 He used satellite DNA as the basis of DNA fingerprinting that shows very high degree of
polymorphism. It was called as Variable Number Tandem Repeats.(VNTR)
 Different steps of DNA fingerprinting are:-
o Isolation of DNA.
o Digestion of DNA by restriction endonucleases.
o Separation of DNA fragments by gel electrophoresis.
o Transferring (blotting) of separated DNA fragments to synthetic membranes,
such as nitrocellulose or nylon.
o Double stranded DNA made single stranded.
o Hybridization using labeled VNTR probe.
o Detection of hybridized DNA fragments by autoradiography.
 The VNTR belongs to a class of satellite DNA referred to as mini-satellite.
 The size of VNTR varies from 0.1 to 20 kb.
 After hybridization with VNTR probe the autoradiogram gives many bands of different
sizes. These bands give a characteristic pattern for an individual DNA. It differs from
individual to individual.
 The DNA from a single cell is enough to perform DNA fingerprinting.
Applications:
 Test of paternity.
 Identify the criminals.
 Population diversity determination.
 Determination of genetic diversity.

***********************************************************************************************

54
 Chapter 7- EVOLUTION
Evolution is a process that results in heritable changes in population spread
over many generations leading to diversity of organisms on earth.
ORIGIN OF LIFE:

The Big Bang theory:

The Big Bang theory states that a huge explosion occurred, the universe expanded, temperature
came down and hydrogen and helium were formed later. The galaxies were then formed due to
condensation of gases under gravitation. A singular huge explosion unimaginable in physical
term. In the solar system of the Milky Way galaxy, earth was supposed to have been formed
about 4.5 billion years back.

Conditions prevailed in the primitive atmosphere:

* There was no atmosphere on primitive earth.
* Water vapor, methane, carbon dioxide and ammonia released from molten mass covered the surface.
* The UV rays from the sun broke up water into Hydrogen and oxygen and lighter H2 escaped.
* Oxygen combined with ammonia and methane to form water, CO2 and others.
* The ozone layer was formed.
* As it cooled, the water vapor fell as rain, to fill all the depressions and form oceans.
* Life appeared 500 Million years after the formation of earth.
Theories of Origin of life:
(i) Theory of special creation states that God created life by his divine act of creation.
(ii) Theory of panspermia/cosmozoic theory, given by early Greek thinkers states that the spores or
panspermia came from outer space and developed into living forms.
(iii) Theory of spontaneous generation states that life originated from decaying and rotting matter like
straw, mud, etc. (a) Louis Pasteur rejected the theory of spontaneous generation and demonstrated
that life came from pre-existing life. (b) In his experiment, he kept killed yeast cells in pre-
sterilised flask and another flask open into air. The life did not evolved in the former but new living
organisms evolved in the second flask.
(iv) Theory of chemical evolution or Oparin-Haldane theory states that life originated from pre-existing
non-living organic molecules and that formation of life was preceded by chemical evolution. The
conditions on the earth that favoured chemical evolution were very high temperature, volcanic
storms and reducing atmosphere that contained CH4,NH3, water vapour, etc.
Urey and Miller experiment:

In 1953, S.L. Miller an American Scientist created similar primitive earth conditions in a
laboratory and simulated the primitive earth.
* The conditions created for simulating the primitive erath were - High temperature, Volcanic
storms and Reducing atmosphere containing CH4, NH3 etc.
* He created electric discharge in a closed flask to raise temperature upto 800oC as it was in
primitive earth.
* Used CH4 H2, NH3 and water vapor inside the flask.
* He observed the formation of amino acids.

55
 * Miller observed the synthesis of amino acids from simple inorganic chemicals in simulated
condition in the laboratory.
* In similar experiments others observed, formation of sugars, nitrogen bases, pigment and fats.
* Analysis of meteorite content also revealed similar compounds indicating that similar processes are
occurring elsewhere in space.

Theory of biogenesis:
* The first non-cellular forms of life could have originated 3 billion years back.
* They would have been giant molecules (RNA, proteins, Polysaccharides, etc).
* These capsules reproduced their molecules perhaps, named as coaservates.
* The first cellular form of life did not possibly originate till about 2000 millions years ago.
* The first cellular forms of life were probably unicellular.
* All life forms were in water environment only.
* This theory of biogenesis from non-living molecules was accepted by majority.
EVOLUTION OF LIFE FORMS – A THEORY:
 Conventional religious literature tells us about the theory of special creation.
 The theory of special creation has three connotations:-
o All the living organisms (species types) that we see today were created as such.
o The diversity was always the same since creation and will be same in future.
o Earth is about 4000 years old.
Challenge to special creation theory:
 Observation made during a sea voyage in a sail ship called H.M.S. Beagle round the world.
Charles Darwin concluded that existing life forms share similarities to varying degrees not
only among themselves but also with life forms that millions of years ago.
 Many such life forms exist any more. There had been extinctions of different life forms in
the years gone by just as new forms of life arose at different periods of history of earth.
 There has been gradual evolution of life forms.
 Any population has built in variation in characteristics.

56
  Those characteristics which enable some to survive better in natural conditions (climate,
food, physical factors, etc) would outbreed others that are less-endowed to survive under
such natural condition.
 Survival of the fittest. The fitness according to Darwin refers ultimately and only leaves
more progeny than others.
 These, therefore, will survive more and hence are selected by nature. He called it as natural
selection.
 Alfred Wallace, a naturalist who worked in Malay Archipelago had also come to similar
conclusions around the same time.
The geological history of earth closely correlates with the biological history of earth.\

EVIDENCES FOR EVOLUTION

Paleontological evidence:
* Fossils are remained of hard parts of life-forms found in rocks.
* Different-aged rock sediments contain fossils of different life-forms who probably died during the
formation of the particular sediment.
* They represent the extinct organisms (e.g. Dinosaurs).
* A study of fossils in different sedimentary layers indicates the geological period in which they
existed.
* The study showed that life-forms varied over time and certain life forms are restricted to certain
geological time-span.
* Hence new lives have arisen at different times in the history of earth.
* All this called Paleontological evidence.
Comparative anatomy and morphological evidence:
* Comparative anatomy and morphology shows similarities and differences among organisms of
today and those that existed years ago.
Divergent evolution:
* Whale, bats, cheetah and human share similarities in the pattern of bones of forelimbs.
* These forelimbs perform different functions in these animals, they have similar anatomical structure
– all of them have humerus, radius, ulna, carpals, metacarpals and phalanges in their forelimbs.
* Hence in these animals, the same structure developed along different directions due to adaptation to
different needs.
* This is divergent evolution and these structures are homologous.
* Homology indicates common ancestry.
* Other examples of homologous organ are vertebrate hearts and brains.
* Thorn of Bougainvilleaand tendrils of Cucurbitarepresent homology.
Convergent evolution:
* Wings of butterfly and of birds look alike.
* They are anatomically similar structure though they perform similar function.
* Hence analogous structures are a result of convergent evolution.
* Eye of octopus and eye of mammals.
* Flippers of Penguins and Dolphins.
* Sweat potato (root modification) and potato (stem modification).
Biochemical evidences:
* Similarities in proteins and genes performing a given function among diverse organisms give clues
to common ancestry.
Embryological support for evolution:
* Proposed by Ernst Heckel based upon observation of certain features during embryonic stage
common to all vertebrates that are absent in adult.
57
 * The embryos of all vertebrates including human develop a row of vestigial gill slits just behind the
head but it is a functional organ only in fish and not found in any other adult vertebrates.
* This is disproved on careful study performed by Karl Ernst von Baer. He noted that embryos
never pass through the adult stages of other animals.
Evolution by natural selection:
* Based on observation of moth population in England made in 1850.
* Before industrialization set in, it was observed that there were more white-winged moths on trees
than dark-winged or melanised moths.
* After industrialization i.e. 1920 there were more dark-winged moths in the same area i.e. the
proportion was reversed.
Evolution by anthropogenic action:
* Excess use of herbicides, pesticides etc., has only resulted in selection of resistant varieties in a
much lesser time scale.
* This is also true for microbes against which we employ antibiotics or drugs against eukaryotic
organisms/cell.
* Hence resistance organisms/cells are appearing in a time scale of months or years and not in
centuries.
* These are the examples of evolution by anthropogenic action.
* Evolution is a stochastic process based on chance events in nature and chance mutation in the
organisms.

ADAPTIVE RADIATION

Darwin’s Finches:
Link for the Video of Adaptive Radiation:
https://www.youtube.com/embed/rMCP2n7VXH8

* In Galapagos Islands Darwin observed small black birds later called Darwin’s Finches.
* He realized that there were many varieties of finches in the same island.
* All the varieties, he came across, evolved on the island itself.
* Form the original seed-eating features, many other forms with altered beaks arose, enabling them to
become insectivorous and vegetarian finches
* This process of evolution of different species in a given geographical area starting from a point and
literally radiating to other areas of geography (habitats) is called adaptive radiation.

58
Australian marsupial:
* A number of marsupials each different from the other evolved from an ancestral stock. But all
within the Australian island continent.
* When more than one adaptive radiation appeared to have occurred in an isolated geographical area
(representing different habitats), one can call this convergent evolution.
* Placental mammals in Australia also exhibit adaptive radiation in evolving into varieties of such
placental mammals each of which appears to be ‘similar’ to a corresponding marsupial (e.g.
placental wolf and Tasmanian wolf-marsupial).
BIOLOGICAL EVOLUTION:
* The essence of Darwinian Theory about evolution is natural selection.
* The rate of appearance of new forms is linked to the life cycle or the life span.
* There must be a genetic basis for getting selected and to evolve.
* Some organisms are better adapted to survive in an otherwise hostile environment.
* Adaptive ability is inherited.
* It has genetic basis.
* Fitness is the end result of the ability to adapt and get selected by nature.
* Branching descent and natural selection are the two key concepts of Darwinian Theory of
Evolution.
Lamark theory of evolution: (theory of inheritance of acquired characters)
* French Naturalist Lamark had said that evolution of life forms had occurred but driven by use and
disuse of organs.
* He gave the example of Giraffes who in an attempt to forage leaves on tall trees had to adapt by
elongation of their necks.
* They passed on this acquired character of elongated neck to succeeding generations.
* Giraffes, slowly over the years, came to acquire long necks.

MECHANISM OF EVOLUTION:
* In the first decade of twentieth century, Hugo deVries based on his work on evening primrose
brought fourth the idea of mutations.
* Mutation is the large difference arising suddenly in a population.
How deVries theory of mutation differs from Darwin’s theory of natural selection?
* It is the mutation which causes evolution and not the minor variations that Darwin talked about.
* Mutations are random and directionless while Darwinian variations are small and directional.
* Evolution for Darwin was gradual while deVries believed mutation caused speciation and hence
called it saltation (single step large mutation).
HARDY – WEINBERG PRINCIPLE:
* In a given population one can find out the frequency of occurrence of alleles of a gene on a locus.
* This frequency is supposed to remain fixed and even remain the same through generations.
* Hardy-Weinberg principle stated it using algebraic equations.
* The principle states that allele frequencies in a population are stable and is constant from generation
to generation.
* The gene pool (total genes and their alleles in a population) remains a constant. This is called
genetic equilibrium:
* Sum total of all the allelic frequencies is 1.
2 2 2
* (p + q) = p + 2pq + q = 1.
* When frequency measured, differs from expected values, the difference (direction) indicates the
extent of evolutionary change.
* Disturbance in genetic equilibrium, or i.e. change of frequency of alleles in a population would then
be interpreted as resulting in evolution.

59
 * Five factors are known to affect Hardy-Weinberg equilibrium:
i)Gene migration or gene flow.
ii)Genetic drift.
iii) Mutation.
iv) Genetic recombination.
v)Natural selection.

* Gene migration: When migrations of a section of population to another place occur, gene
frequencies change in the original as well as in the new population. New genes /alleles are added to
the new population and these are lost from the old population.
* Gene flow: Gene migration occurs many time is termed as gene flow.
* Genetic drift: change in gene frequency takes place by chance.
* Founder effect: sometimes the change in allelic frequency is so different in the new sample of
population that they became a different species. The original drifted population becomes founder
species and the effect is called founder effect.
Operation of natural selection on different trait:
 Natural selection can lead to :
o Stabilization: in which more individuals acquire mean character value.
o Directional changes i.e. more individuals acquire value other than the mean character value.
o Disruption: more individuals acquire peripheral character value at both ends of the distribution
curve.
ORIGIN AND EVOLUTION OF MAN:
* About 15 mya primates called Dryopithecus and Ramapithecus were existing.
* They were hairy and walked like gorillas and chimpanzees.
* Ramapithecus was more man like while Dryopithecus was more ape-like.
* Few fossils of man-like bones have been discovered in Ethiopia and Tanzania.
* Two mya Australopithecines probably lived in East African grasslands.
* The first human-like being the hominid and was called Homo habilis.
* Fossils discovered in Java in 1891 revealed the next stage i.e. Homo erectus about 1.5 mya.
* Neanderthal man:
* Homo sapiens: Arose in Africa and moved across continents and developed distinct races.

Evolution video:
https://www.youtube.com/embed/RTX9si5RBb0

60
 Chapter 8 - HUMAN HEALTH AND DISEASE
HEALTH :- Health is defined as the state of complete physical , mental and social well being.
DISEASE :- Disease is the condition when the functioning of one or more organs or systems of the body is
/ are adversely affected
* Factors important to maintain good health:-
Good Health can be attained by the following :-
* Balanced Diet
* Personal Hygiene
* Regular Exercise
* Good Habit
* Awareness about diseases
* Vaccination
* Proper waste disposal
* Control of vectors
* Consumption of hygienic ( clean ) food and water
Types of Diseases

SL COMMUNICABLE/INFECTIOUS NON COMMUNICABLE / NON INFECTIOUS

DISEASES DISEASES

1 These are the diseases that are easily These are the diseases that are not
transferred from diseased to a transferred from diseased to a healthy
healthy person person

2 They are caused by pathogens eg. They are caused due to genetic factors,
Bacteria,Viruses,Fungi, Helminthes , nutritional deficiencies, unhealthy life
Protozoa etc styles etc.

Some common Infectious Diseases :

* BACTERISEASES :- Pneumonia,T.B. , Cholera , Typhoid etc.
* VIRAL DISEASES :- Common Cold, AIDS ,Chikungunya ,Dengue,
Hepatitis , Polio , Measles
* FUNGAL DISEASES :- Ring worm
* PROTOZOAL DISEASES :- Amoebiasis , Malaria
* HELMINTHIC DISEASES :- Ascariasis , Filariasis , Taeniasis
* VIRAL DISEASES :-

61
 COMMON COLD :- Causal Organism :- Rhinoviruses
Organs Affected :- Nasal & Respiratory passages
Spreads By :-
i. Droplets released during cough & sneezing by an infected person.
ii. Contaminated objects / articles.
Symptoms :-
i. Nasal congestion & discharge
ii. Sore throat
iii. Cough
iv. Tiredness
v. Headache

62
vi. Hoarseness
* DENGUE
* Causal Organism :- Dengue Virus
Organs Affected :-
Spreads By :- Bite of Aedes aegypti mosquito.
Symptoms :-
i. Fever
ii. Headache
iii. Muscle & joint Pain
iv. Characteristic skin rash (like measles )
v. In severe cases it becomes haemorrhagic fever , where the platelet count of blood decreases
Prevention :- By controlling the vector mosquito.
CHIKUNGUNYA
* Causal Organism :-Alpha Virus having ss RNA as its genome
Organs Affected :-
Spreads By :- Bite of Aedes aegypti mosquito.
Symptoms :-
i. High fever
ii. Rashes on the trunk
iii. Nausea
iv. Vomiting
v. Multiple joint pain
Prevention & Control :-
Mosquito control is the effective prevention method.
TYPHOID
* Causal Organism :- Salmonella typhi (Bacteria )
Organs Affected :- Small Intestine . Intestinal perforations leading to death may occur in severe
cases.
Spreads By :- Contaminated food and water
Symptoms :-
i. Sustained high fever ( 103 – 104 o F )
ii. Stomach Pain
iii. Loss of appetite
iv. Constipation
v. Headache
How to Confirm :- Widal Test
PNEUMONIA
* Causal Organism :- i) Streptococcus pneumoniae & ii) Haemophilus enfluenzae
Organs Affected :- Lung Alveoli . Alveoli become filled with a fluid , resulting in severe difficulty
in breathing / respiration.
Spreads By :-
i. Droplets from an infected person
ii. Sharing the contaminated articles

63
 Symptoms :-
i. Fever
ii. Headache
iii. Cough
iv. Chills
v. In severe cases , the lips and fingernails may turn greyish
MALARIA ( a protozoal disease )
Causal Organism :- Plasmodium species
* P. malariae
* P. vivax
* P. falciparum ( Malignant malaria )
Organs Affected :-
Spreads By :- Bite of Female Anopheles mosquito that transfers the
Sporozoits (Infectious stage ) of Plasmodium.
Symptoms :-
i. High fever and chill
Prevention & Control :-
Disease can be controlled by eradicating the mosquitoes & avoiding mosquito bite by using
mosquito repellents , mosquito nets etc.
Treatment :- Use of Chloroquins.
LIFE CYCLE OF MALARIAL PARASITE :-
1. The sporozoites enter the body at the bite of mosquito, reach the liver through blood and multiply
within the liver cells.
2. Such liver cells burst and release the parasites into blood.
3. In the blood the parasite attacks RBCs , multiply asexually and cause their rupture. The rupture of
RBC releases a toxin called Haemozoin , which is responsible for the recurring High Fever and the
Chill
4. Sexual stages (Gametocytes) develop in the red blood cells.
5. The parasite then enters the female anopheles mosquito along with the blood when it bites the
infected person.
6. Further development occurs in the stomach wall of the mosquito
7. The gametes fuse to form a zygote .
8. The zygote undergoes further development in the body of the mosquito to form sporozoites.
9. The Sporozoites are transported to and stored in the Salivary gland of the mosquitoes and are
transferred to the human body during the bite of the mosquito.

Common Diseases:
https://www.youtube.com/embed/YA9KiI7gW5Q

64
PPT for Common Human Diseases :
https://docs.google.com/presentation/d/1qznWGDl_9nFvPzjrV-
8o2Vv4giwvDtK_/edit?usp=sharing&ouid=107171444976724466844&rtpof=true&sd=true

AMOEBIC DYSENTRY (Protozoal Disease )

* Causal organism :- Entamoeba histolytica

* Mode of Dispersal :- Through contaminated food and water.
* House flies act as mechanical carriers and transfer the parasite from the faeces of infected person to
the food articles and water.
* Organ Affected : The parasite resides in the large intestine.
* Symptoms :
* i) Abdominal pain and cramps.
* Stool with excess mucus and blood clots.
* Constipation alternating with diarrhoea.

65
 RING WORM ( FUNGAL DISEASE)
* Causal Organism : Microsporum , Epidermophyton , and Trichophyton.
* Symptoms :
* i) Dry scaly lesions on skin , nails , and scalp.
* Ii) Lesions are accompanied by itching.
Ringworms are generally acquired from soil or by direct contact.
ASCARIASIS (HELMINTHIC DISEASE)
Causal Organism :- Ascaris lumbricoides (Round Worm )
Symptoms :-
i) Blockage oftheintestinal passage
ii) Anaemia
iii) Abdominal / Muscular pain
iv) Internal bleeding
v) Nausea and headache
Mode of Dispersal :
Through contaminated food and water as eggs of parasite excreted by the infected person
contaminate soil, plants and water
FILARIASIS (ELEPHANTIASIS) Protozoal Disease
* Causal Organism : Wuchereria bancrofti and W. malayi wer limbs causing them to swell like that of
an ( Filarial Worm)
* Mode of Dispersal : Through the bite of Female Culex Mosquito
* Symptoms :-
* i) They normally cause inflammation of the organs in which they live for many years.
* ii) They normally affect the lymph vessels , of the lower limbs, causing them to swell like that of an
Elephant , hence called Elephantiasis.
* iii) Genital Organs may also be affected leading to gross deformation.
PREVENTION & CONTROL OF INFECTIOUS DISEASES :-
* i) Maintenance of personal hygiene.
* Maintenance of Public hygiene.
* Eradication of vectors and their breeding places.
* Vaccination and immunisation for disease like Polio , Diphtheria, tetanus etc.
* Use of antibiotics and drugs to treat the infected person.

AIDS : ACQUIRED IMMUNODEFICIENCY SYNDROME

* Causal Organism :HIV(Human Immunodeficiency Virus) a retrovirus
* Transmission of HIV occurs in one or more of the following ways :
* i) By sexual contact with the infected person .
* ii) Transfusion of the contaminated blood and blood products.
* iii)Sharing of infected needles.
* iv) From infected mother to the developing baby (inside uterus) through placenta.
* Following individuals are at high risk of getting the disease :
* i) Those who have multiple sexual partners.
* ii) Drug addicts taking the drugs intravenously.

66
* iii) Individuals who require repeated blood transfusion.
* iv) Children born to an infected woman.
* LIFE CYCLE OF HIV :-
* i) The Virus after getting into the body of a person , enters the Macrophages.
* ii) The RNA replicates and DNA is formed by Reverse Transcriptase.
* The viral DNA gets incorporated with the host cell DNA and directs the infected cell to produce the
viral particles.
* The Macrophages continue to produce Virus particles
* The virus then enters the helper T-Lymphocytes ( TH), replicates and forms the progeny viruses.
* The progeny viruses released in the blood attack other helper T- Lymphocytes and there is a
progressive decrease in the number of helper T-Lymphocytes in the body of the infected persons.
* The person becomes easily infected by bacteria like Mycobacterium , viruses and even parasites
like Toxoplasma.
* The person is unable to protect himself / herself against any infection.

* Prevention of AIDS :

67
 * NACO ( National AIDS Control Organisation ) and Non-governmental organisations are trying
their best to educate people about AIDS.

* WHO (World Health Organisation ) has started a number of programmes to prevent spreading of
HIV infection. Some such steps are :-

* i) Ensuring use of disposable needles & syringes.

* ii) Checking blood forHIV
* iii) Free distribution of condomes and advocating safe sex.
* iv)Controlling drug abuse.
* v) Promoting regular check-up for HIV susceptible populations etc.
* DIAGNOSIS OF AIDS :
* By ELISA Test :( Enzyme – Linked Immuno Sorbant Assay )

TREATMENT : Treatment with anti-retroviral drugs is only partially effective. They can only prolong the
life of the patient but cannot prevent death.

CANCER :- GENERAL ACCOUNT

* Transformation of normal cells into cancerous cells is induced by carcinogens.
* Carcinogens are those physical, chemical , and biological agents which bring about uncontrolled
proliferation of cells (Cancer )
Types of Carcinogens :
* i)Physical Carcinogens :Eg. U.V. Rays,X – rays , gamma rays.
* ii) Chemical Carcinogens : Eg. Aniline dyes,chemicals present in tobacco smoke.
* iii) Tumour viruses (Oncogenic Viruses)
* Difference between cancerous cells and Normal Cells )
* i) There is breakdown of the regulatory mechanism which control normal cells growth , cell
division , and differentiation.
* ii) Cancer cells do not show contact inhibition and show uncontrolled cell divisions.
* iii) Cancer cells show Metastasis i.e. they detach from the tumours and move to distant sites
through body fluids and develop secondary tumours.

TYPES OF TUMOURS :
i) Benign Tumours :- They remain confined to their original location and do not spread to other parts.
They cause little damage.
ii) Malignant Tumours :-
a) They are masses of neoplastic / proliferating cells which grow rapidly , invade and damage the
surrounding normal tissue/cells.
b) These cells compete with the normal cells for vital nutrients and disrupt the normal metabolism.
c) These cells show the property of Metastasis.
DIAGNOSIS OF CANCER
i) Biopsy and Histopathological studies of the tissue.
ii) Blood and Bone Marrow Tests for increased cell counts as in Leukaemia.
iii) Use of techniques like Radiography, MRI (Magnetic Resonance Imaging) and CT – Scan (
Computed Tomography ) for cancer of internal organs.
iv) Use of Antibodies against cancer specific antigens.

68
 v) Applying principles of Molecular Biology to detect gene in individuals with inherited
susceptibility to certain cancers.
TREATMENT OF CANCER
i) SURGERY :- The Tumour cells are surgically removed to reduce the load of cancerous cells.
ii) RADIOTHERAPY :- The Tumour cells are irradiated wholly but taking care of the surrounding
normal
cells.
iii) CHEMOTHERAPY :- Certain drugs are used to kill the cancerous cells , but majority of the
drugs have side effects like hair loss. Anaemia, etc.
iv) IMMUNOTHERAPY :- This involves the use of biological response modifiers like alpha
interferon , which activate the immune system and help in destroying the tumour.

IMMUNE SYSTEM

Definition of Immunity :- It refers to the overall ability of a living body to fight against diseases.

69
 IMMUNITY
Ability of the body's immune system to protect the body from disease

ACQUIRED IMMUNITY
INNATE IMMUNITY
CHARACTERSTICS FEATURES
ACTIVE PASSIVE
1. NON SPECIFIC
2. PRESENT AT THE TIME OF BIRTH
CHARACTERSTICS FEATURES
1. SPECIFICITY
2. DIVERSITY
TYPES OF BARRIERS 3. DISTINGUISH BE
TWEEN SELF NON SELF
1. PHYSICAL BARRIER - SKIN
4.MEMORY
2. PHYSIOLOGICAL BARRIER - HCL & SALIVA
5. ACQUIRED DURING LIFETIME
3. CELLULAR BARRIER - W.B.C ,MACROPHAGE
4. CYTOKINE BARRIER - INTERFERON
CELLS OF AQUIRED IMMUNITY
(LYMPHOCYTE)

T-LYMPHOCYTES
B-LYMPHOCYTE
1. HELP B-CELLS TO PRODUCE
1.PRODUCE ANTIBODY IN
ANTIBODY RESPONSE TO
2.DIRECTLY ATTACKS PATHOGEN (ANTIGEN)
PATHOGENS
3.RETAINS MEMORY

INNATE IMMUNITY :- It refers to all the defense elements with which an individual is born and is
always available to protect the body. It is nonspecific and of four different types.

1. PHYSICAL BARIERS :
i) The Skin - main barrier : (Surface Area=1.73 m2)

ii) The mucus-coated epithelium lining of the

a) Respiratory,

70
 b) Gastro-intestinal,
c) Urogenital tracts,
where mucus helps in trapping the microbes.

2. PHYSIOLOGICAL BARRIERS: They prevent microbial growth : Examples:

3. CELLULAR BARRIER : They include the :

A. Leucocytes (WBC) which phagocytose and destroy the microbes.

i) PMNL - Neutrophils
ii) Monocytes
iii) Lymphocytes (Natural killers)

B. Macrophases (in the Tissues)

4. CYTOKINE BARRIER :

Interferon (Proteins) produced by viral infected cells & protect the non-infected cells from further
Viral infections.
71
 ACQUIRED IMMUNITY: It refers to the immunity a person acquires after birth, either by
contracting the disease or by vaccination.

Characteristic features of acquired immunity:

i) SPECIFICITY: It is pathogen specific.
ii) MEMORY: The memory cells produced during the first encounter with the pathogens evoke a
heightened immune response in the further encounters.
iii) Distinguishes between self and non self.

Types of Immunity Link for the video:

https://www.youtube.com/embed/odIdD_uwIPE

Immunity - Presentation:
https://docs.google.com/presentation/d/1DdZLgJ1nNNyHY4ek9OmdvIn5q0AVDt6E/edit?
usp=sharing&ouid=107171444976724466844&rtpof=true&sd=true

TYPES OF IMMUNE RESPONSES:-

1. PRIMARY IMMUNE RESPONSE :- An Immune Response of low Intensity, when our body
encounters a pathogen for the first time

2. SECONDARY IMMUNE RESPONSE :- (Anamnestic Response) A highly intensified Immune

Response when our body encounters the same pathogen for the second time. Why ? Because our body
keeps a memory of the pathogen when encoun- tered it for the first time.

TYPES OF ACQUIRED IMMUNITY:

A) HUMORAL or ANTIBODY MEDIATED IMMUNITY :-

i) It consists of the antibodies, that are circulating in the body fluids (Humors)

ii) The antibodies produced by B- Lympho- cytes in response to the antigens are collectively called
Immunoglobins and are of various types like IgA,IgD,IgE, IgG,IgM .

ANTIBODY MOLECULE (Immunoglobins):

i) Each antibody consists of four polypeptide chains, held

together in the form of Y where the tips of two upper
arms bind to the antigens in a lock and key manner to
form antigen-antibody complex.

ii) The two polypeptide chains are long and called

heavy(H) chains , while the other two are short and
called light (L) chains. Hence, the antibody is referred to

72
 as H2L2.

B) CELL MEDIATED IMMUNITY : CMI

It is mediated by T-Lymphocytes. There are two groups of T-Lymphocytes
i) Cytotoxic / Killer T-Cells – which kill the specific target cell by a variety of mechanisms.
ii) Helper T – cells : which activate the specific B-cells to produce antibodies.
T-Lymphocytes are responsible for graft rejection.

On the basis of Involvement of body in the production of Antibodies, immunity is of two types :
a) Active Immunity and b) Passive Immunity.

Sl Active Immunity Passive Immunity

No.
i) When the antibodies are developed When antibodies developed in other
by our own cells in response to the vertebrates in response to deliberate injection
antigens, it is called active immunity of antigen , are injected into our body, it
constitutes passive immunity.
ii) It takes time to develop It shows fast response hence it is used when
immunity(slow response) immune response has to be faster. E.g. in case
of snake bite.
iii) It stays in body for longer period. It stays in body for short period.
Example : Example :
a) Immunity developed during a) Immunity given to the infant by antibodies
natural in colostrum
exposure to pathogens b) Immunity given by Tetanus Antitoxin
b) Immunity developed by
vaccination.
VACCINATION :-

Vaccination is the process of introducing a preparation of antigenic proteins of pathogens or killed or

inactivated /attenuated pathogens into the body, to generate the immune response / antibodies.

ALLERGY :- It refers to the exaggerated / hypersensitive response of the immune system to certain
antigens in the environment.
ALLERGENS :- The substance / agent which causes the hypersensitive reaction of the immune system
to certain antigens present in the environment.

Example :- Dust , Mites , Pollen Grains , Animal Dander etc.

The antibodies produced in response to allergens are Ig E type.

COMMON SYMPTOMS OF ALLERGY :-

Under the influence of allergens certain chemicals like Histamine and Serotonin are released from mast
cells. These chemicals induce the symptoms of allergy.

i) Sneezing (ii) Watery eyes (iii) Rashes (iv) Running nose (v) Difficulty in breathing.

73
 CAUSES OF ALLERGY: Somehow, modern - day life style has resulted in lowering of immunity
and more sensitivity to allergens. More & more children in metro cities of India suffer from
allergies and asthma due to sensitivity to the environment.

Allergy is due to the release of chemicals like histamine and serotonin from the mast cells.

AUTOIMMUNITY :

Autoimmunity or autoimmune disorders are those disorders which are caused when the body’s
immune system goes off the track and start destroying ‘Self Cells’ and molecules.

Example : Hashimoto’s Thyroiditis, Systemic

Lupus, Rheumatoid Arthritis etc.

MAIN FUNCIONS of IMMUNE SYSTEM

To recognize the foreign molecules(Antigens),

respond to them and keep a memory of them.

COMPONENTS OF IMMUNE SYSTEM :-

1. Lymphoid Organs
2. Lymphoid Tissues
3. Lymphoid Cells
4. Soluble Molecules like Antibodies.
FUNCTIONS OF PRIMARY LYMPHOID
ORGANS :-
1. Bone Marrow :- Here all kinds of blood cells are produced
2. Thymus :- Provides a micro environment for the development and maturation of lymphocytes.

FUNCTIONS OF SECONDARY LYMPHOID ORGANS :

1. SPLEEN :-
i) It mainly contains lymphocytes and phagocytes.
ii) It acts as filter of blood by trapping blood borne microbes
iii) It also acts as a reservoir of Erythrocytes (RBCs)
2. LYMPH NODES :- They act as filters and trap the microbes that have entered the lymph.

3. MALT :- Mucosal Associated Lymphoid Tissues:- Lymphoid tissue located within the mucosal
lining of the major tracts (respiratory , digestive, urogenital tracts) is called MALT.

Different types of lymphoid organs and their functions is being summarized by the help of
following flow diagrams

74
*********************************************

75
 Chapter 9 - MICROBES IN HUMAN WELFARE
A. MICROBES IN HOUSEHOLD PRODUCTS
SL NAME OF FOOD ITEM MICROBE INVOLVED
NO
01 CURD LACTOBACILLUS & other LAB
(Lactic Acid bacteria)
02 DOUGH ( For Idli & Fermented by bacteria. Dough rises due to CO2 produced
Dosa) during fermentation.
03 DOUGH (For making Saccharomyces cerevisiae (Backers Yeast )
Bread)
04 CHEESE
a) Swiss Cheese has large Propionibacterium sharmanii.
holes due to production of
CO2)
b) Roquefort Cheese Ripened by growing a specific fungus in it.

B. MICROBES IN INDUSTRIAL PRODUCTS

SLN NAME OF PRODUCT MICROBE INVOLVED
O
1 BEEVERAGES :-
a)ETHANOL :- (By fermentation Sacharomyces cerevisiae ( Yeast )
of fruit juices & malted cereals)

Remark :- Type of alcoholic drink varies with the type of Raw material & nature of
processing.
a) With Distillation :- Whisky , Brandy & Rum.
b) Without Distillation :- Beer & Wine.
2 ANTIBIOTICS
a)Penicillin :- Penicilium notatum ( Fungi )
Remark :- Penicillin discovered by Alexander Fleming, while working on Staphylococci
bacteria, but it was established as an effective drug by Ernst Chain & Howard Florey.
The Three were awarded Nobel prize in medicine in 1945.
3 ORGANIC ACIDS
a) Citric Acid Aspergillus niger ( Fungus)
b) Acetic Acid Acetobacter aceti ( Bacterium )
c) Butyric Acid Clostridium butylicum ( Bacterium )
d) Lactic Acid Lactobacillus delbrueckii (Bacterium)
4 ENZYMES
a)Lipases Used in detergent formulations.
b)Proteases & Used to clear fruit juices.
c)Pectinases
d)Streptokinases A ‘clot buster’ produced by genetically modified
Streptococcus.

76
 Remark :-Streptokinase is used for removing blood clots in blood vessels of patients suffering
from Myocardial Infractions.
5 BIOACTIVE MOLECULES
a)Cyclosporin A Trichoderma polysporum. ( Fungus )
(Used as Immuno suppresant)
b)Statins (Used for lowering blood Monascus purpureous (Yeast )
cholesterol)
6 BIOGAS PRODUCTION Methanogens like Methanobacterium
Remark :- Methanogens produce Methane,CO2,H2 & H2S by acting over cellulosic compounds
present in cattle dung. They are found in i) Anaeribic sludge, ii)Rumen of cattle, iii) Flooded
Rice Fields, & iv) Marshy places.
7 MICROBES AS BIOCONTROL It refers to the use of controlling pests that relies
AGENTS on natural predators
a)Bacillus thuringiensis ---------- A bacterium whose spores are toxic to certain
insect larvae (butterfly caterpillars) & kill them.
A fungus(free living in soil &root ecosystems) is
b)Trichoderma -------------- effective against several plant pathogens.

They attack insects and other arthropods. These

c)Baculoviruses viruses are excellent candidates for species
(Genus : Nucleopolyhedroviruses) specific, narrow spectrum insecticidal
applications, Useful in IPM in ecologically
sensitive area

Used to get rid of Aphids

c)Ladybird (a beetle with red &
black markings)
Used to get rid of Mosquitoes
d)Dragonfly ------------------------
8 MICROBES AS They fix atmospheric Nitrogen, Organic matter &
BIOFERTILIZERS other soil nutrients.
a BACTERIA
(Fix Atmospheric N2)
i)Symbiotic Bacteria ---------- Rhizobium.
ii)Free Living bacteria ------- Azotobacter & Azospirillum.
b CYANOBACTERIA Nostoc,oscillatoria,Anabaena,Aulosira.
c FUNGI/MYCORRHIZAE Symbiotic association of fungi with the roots of
higher plants. e.g. Glomus
Remark :- Mycorrhizae are beneficial in the following manner :-
i)They absorb phosphorus & passes it on to the plant.
ii)Provide resistance to root borne pathogens.
iii)Tolerance to salinity.
iv)Overall increase in the plant growth & development.

Microbes in Human welfare:

https://www.youtube.com/embed/65sh_0kBuM8

77
SEWAGE TREATMENT :- Municipal waste water is called sewage which may contain large
amount of organic matter & microbes( which may be Pathogenic)Therefore Sewage is Treated in
Sewage Treatment Plant before it is released in the water bodies.
Sewage treatment consists of two steps :-
I. PRIMARY TREATMENT :- Physical process to remove insoluble large & small particles
through Filtration & Sedimentation.
Filtration :- Sewage is passed through wire mesh to remove floating insoluble objects like
polythene.
Sedimentation :- Sewage is then passed into grit chamber, where the grit(soil & small peebles
)are removed by sedimentation.
All the solids that settled , form the Primary Sludge and the supernatant forms the
effluent which is taken to another tank for secondary treatment.
II. SECONDRAY TREATMENT :- ( Biological Treatment ) Two step treatment.
AEROBIC TREATMENT :-
The primary effluent is passed into large aeration tank where it is constantly agitated
mechanically and air is pumped into it. This allows vigorous growth of useful microbes into
flocs ( Flocs = Mass of bacteria + fungal hyphae )
While growing , these microbes consume the major part of the organic matter in the
effluent. This significantly reduces the biochemical oxygen demand(BOD)

A) ANAEROBIC TREATMENT :-
i) The effluent is then passed into a tank where the bacterial flocs are allowed to
sediment. This sediment is called activated sludge.
ii) A small part of the activated sludge is pumped back into the aeration tank to serve
as the inoculum.The remaining major part of the sludge is pumped into large tanks
called anaerobic sludge digestors. Here, anaerobic bacteria grow and digest the sludge.
During this digestion bacteria produce a mixture of gases called Biogas (Methane-
75%,CO2,H2 & H2S)

BIOGAS PLANT :- Its technology was developed

in India due to the efforts of IARI & KVIC.
Models of Biogas plant :-
a) Fixed Dome Type &
b) Floating Gas Holder Type.
BIOCONTROL AGENTS :- It refers to the use of
controlling pests that relies on natural predators

**********************************************************************
78
 Chapter 11 - BIOTECHNOLOGY: PRINCIPLES AND
PROCESSES

Biotechnology deals with techniques of using live organisms or enzymes from organisms to produce
products and processes useful to humans.

 According to EFB, European Federation of Biotechnology -Biotechnology is defined as ‘The

integration of natural science and organisms, cells, parts thereof, and molecular analogues for
products and services’.
 Current Definition:- It is used in a restricted sense today, to refer to such of those processes which
use genetically modified organisms to achieve the same on a larger scale.
Further many other processes and techniques are also included in the biotechnology :-
i) Test tube baby programme.
ii) Synthesizing a gene and using it.
iii) Developing a DNA vaccine.
iv) Correcting a defective gene(Gene Therapy).

The two core techniques that enabled the birth of modern biotechnology :-

(i) Genetic engineering : Techniques to alter the chemistry of genetic material (DNA and RNA), to
introduce these into host organisms and thus change the phenotype of the host organism.

(ii) Maintenance of sterile (microbial contamination-free) ambience in chemical engineering processes

to enable growth of only the desired microbe/eukaryotic cell in large quantities for the manufacture of
biotechnological products like antibiotics,vaccines, enzymes, etc.

 Three basic steps in genetically modifying an organism —

(i) Identification of DNA with desirable genes;
(ii) Introduction of the identified DNA into the host;
(iii) Maintenance of introduced DNA in the host and transfer of the DNA to its progeny.

 Genetic engineering :-

It involves the alteration of the genetic makeup of cell by deliberate and artificial means. It involves
transfer or replacement of genes to create recombinant DNA.

TOOLS OF GENETIC ENGINEERING:-

1. Enzymes :-

79
 a) Restriction Endonuclease enzyme(Molecular scissors)

b) DNA Polymerase enzyme.

c) DNA Ligases ( Molecular glues)

2. Cloning Vectors: -

Restriction enzymes (R.E.) belong to a larger class of enzymes called nucleases. Each R.E functions by
inspecting the length of a DNA sequence. Once it finds its specific recognition sequence, it will bind to the
DNA and cut each of the two strands of the double helix at specific points in their sugar phosphate
backbone. Each restriction endonuclease enzyme recognizes a specific palindromic sequence in the DNA.

e.g. 5’- GAATTC–3’

3’- CTTAAG- 5’
R.E. cut the DNA strand a little away from the centre of the palindrome sites, but between the same
two bases on the opposite strands. This causes the production of two DNA segments with sticky ends.

https://youtu.be/pMKhOCi7X8w

Restriction enzymes are named in the following manner?

i)The first letter of the name comes from the name of the genus of the source bacteria.
ii)The second two letters come from the species of the prokaryotic cell from which they are
isolated.
iii) The letter R is derived from the name of the strain .
iv) Roman numbers following the names indicate the order in which the enzymes were isolated from
that strain of bacteria. e.g. EcoRI

80
 Diagramatic representation of recombinant DNA technology :-

PROCESSES (STEPS ) OF RECOMBINANT DNA TECHNOLOGY :-

1. Isolation of DNA
2. Fragmentation of DNA by restriction enzymes.
3. Isolation of the desired DNA fragment.
4. Amplification of the gene of interest.
5. Ligation of the DNA fragment into a vector using DNA ligase.
6. Transfer of recombinant DNA into the host.
7. Culturing the host cells on a suitable medium on a large scale.
8. Extraction of the desired product.
9. Down stream processing of the product as a finished product ready for marketing.

rDNA Technology
https://www.youtube.com/embed/xZAD11Sc-a8

81
 rDNA Technology at a glance

Isolation of DNA & cutting of Gel Electrophoresis to

DNA by restriction separate the DNA and PCR (Polymerase Chain
endonuclease enzyme Histone protein Reaction) of desired gene

Down streaming Insertion and ligation of alien

Obtaining of Foreign Gene DNA with Cloning Vector by
Processing product in bioreactor DNA ligase into the host

Separation & isolation, Visualisation &extraction of DNA fragments :-

DNA has to be isolated in pure form for the action of restriction enzymes.
i) DNA can be released from cells by digesting the cell envelop by the use of enzymes like lysozyme
for bacterial cells, chitinase for fungal cells, and cellulase for plant cells.
ii) Since DNA is intertwined with histone proteins and RNA, proteins are removed by proteases and
RNA are removed by ribonucleases.
iii) Other impurities bare removed by employing suitable treatments.
iv) The purified DNA is precipitated by the addition of chilled ethanol. It is seen as fine threads in
suspension.

82
Gel Electrophoresis is the technique of separating DNA segment, in which the negatively charged DNA
segments are forced to move towards anode(+ve electrode) under an electric field through a medium or
matrix.(Commonly Agarose gel)

Visualisation :-The separated DNA fragments can be visualised after staining the DNA by ethidium
bromide followed by exposure to UV rays. A bright orange coloured bands of DNA is observed.

Extraction:- The separated bands of DNA are cut out from the agarose gel & extracted from the gel piece.
This process is called as elution.

CLONING VECTORS :-

 PLASMIDS AND BACTERIOPHASES ARE COMMONLY USED CLONING VECTORS.

 NOWADAYS , GENETICALLY ENGINEERED /SYNTHETIC VECTORS ARE ALSO USED

FOR EASILY LINKING THE FOREIGN DNA AND SELECTION OF RECOMBINANTS
FROM NONRECOMBINANTS.

Making of rDNA at a glance:

Plasmid or Cloning Vector

AmpRGene TetR Gene AmpR gene or tetR gene

Can be cut by Pst I or Can be cut by BamH I and

Insertion fails
Pvu I for the insertion of SalH I for the insertion of
alien DNA Alien DNA

Recombinant DNA Recombinant DNA Recombinant DNA not

produced with the produced by the foreign produced
foreign gene gene

Transformed Bacteria Transformed Bacteria Non transformed Bacteria

83
 FOLLOWING FEATURES ARE REQUIRED TO FACILITATE CLONING IN A VECTOR :-

I) ORIGIN OF REPLICATION :- (ori) This is a sequence of base pairs on DNA where replication
starts. Any piece of DNA linked to this sequence can be made to replicate with in the host cells.
This sequence is also responsible for controlling the copy number of linked DNA.

II) SELECTABLE MARKER :- Marker is a gene which helps in selecting the transformants (those
host cells which contain the gene of interest) and eliminating the non transformants.

Common selectable markers for E.coli include the genes encoding resistance to antibiotics such as
ampicillin, chloramphanicol, tetracycline and canamycin or gene for b-galactosidase which can be
identified by colour reaction

III) CLONING / RECOGNITION SITE :- In order to link the alien DNA , the vector needs to have
very few, preferably single recognition site for the commonly used restriction enzymes.

The ligation of alien DNA is carried out at a restriction site present in one of the two
antibiotic resistant genes. Eg. A foreign DNA is inserted at BAMHI site of tetracycline resistant
gene in the vector PBR322 ( A plasmid).

Some cloning vectors :-

a) Agrobacterium tumefaciens :- It is the bacterium that infects a number of dicot plants and
transfers a piece of its DNA known as tDNA,which transforms the normal plant cells into tumor
cells. Ti plasmid ( Tumor inducing plasmid) of this bacterium has been modified (Disarmed ) and
used as a cloning vector.

b) Retro Viruses :-They infect normal cells and transform them into cancerous cells.Such retroviruses
have been modified (Disarmed ) and used as vectors for transferring DNA into animal cells.

IV) SMALL SIZE OF THE VECTOR :-

Methods of introducing a recombinant DNA into a competent host :-

84
Since DNA is hydrophilic molecule, it cannot pass through cell membranes. Four methods to introduce the
gene of interest into a host.

1. For Bacterial Cell :-(a prokaryotic cell )

a) In order to force bacteria to take up the plasmid, the bacterial cells must first be made ‘competent’ to take
up DNA. This is done by treating them with a specific divalent Cation, eg. Calcium ion, which increases
the efficiency with which DNA enters the bacterium through pores in its cell wall.
b) Recombinant DNA can then be forced into such cells by incubating the cells with recombinant DNA on
ice, followed by placing them briefly at 420C(heat Shock), and then putting them back on ice.This enables
the bacteria to take up the recombinant DNA.

2. MICROINJECTION:- (Suitable for animal cells) In this method , recombinant DNA is directly
injected into the nucleus of an animal cell.

3. BIOLISTICS or GENE GUN :- (Suitable for plants ) In this method plant cells are bombarded with
high velocity microparticles of gold or tungsten coated with DNA.

4. DISARMED PATHOGEN VECTORS :- Such vectors when allowed to infect the cell, transfer the
recombinant DNA into the host.

PCR :- polymerase chain reaction ( Discovered by K Mullis )

It is a technique by which any piece of DNA can be quickly amplified (copied many times ) without using
cells. Each cycle of PCR consists of three steps :-

1. Denaturation of DNA :- Double stranded DNA is denatured by using high temperature.

2. Primer Annealing :-Primers are chemically synthesised oligonucleotides that are complementary to
the regions of DNA segments of interest.
3. Extension of Primers :- Thermostable DNA Polymerase enzyme(Taq Polymerase) extends the
primers using the nucleotides provided in the medium and the genomic DNA as template.

PCR - Link for the video:

https://www.youtube.com/embed/09whdt6PPs0

85
 PCR
Polymerase Chain
Reaction

Denaturation 940c Extension Use of Taq

Annealing Use of
Separation of DNA Polymerase->
Primers Polymerisation
Strands

86
 Introduction of Alien DNA in the competent host at a glance:

0
• Treatment with • Heat (42 c )&
0
divalent cation ice cooling (0 c)
++
Ca ions treatment

Compet
ent Competant
Bacterial
Bacteria Cell
l Cell

Plant Cell
Animal
Cell
• Gene • Micro-injection
Gun/Biolistic method
Method

BIOREACTORS :-

The bioreactors can be thought of as vessels in which raw materials are biologically converted into specific
products by microbes,plants and animal cells and/or their enzymes.

A bioreactor has the following components :-

i) An agitator system.
ii) An oxygen delivery system.
iii) A foam control system.
iv) A temperature control system.
v) pH control system.
vi) Sampling ports.

Downstream processing :-

The products obtained from the genetically modified microorganisms in a biorector is subjected to a series
of processes ( collectively called downstream processing) before it is made into a finished product ready
for marketing.

The two main processes are :-

a) Separation and b) Purification.

The product is then formulated with suitable preservatives. Such formulations have to undergo clinical
trials, in case of drugs.

87
 Chapter 12 - BIOTECHNOLOGY AND ITS
APPLICATIONS
Critical areas of Biotechnology:
• Providing the best catalyst in the form of improved organism usually a microbe or pure enzyme.
• Creating optimal condition through engineering for a catalyst to act.
• Downstream processing technologies to purify the protein/organic compound.

BIOTECHNOLOGICAL APPLICATIONS IN AGRICULTURE:

The focus have been made to increase the food production through Biotechnology in the following areas.
 Agrochemical based agriculture
 Organic Agriculture
 Genetically engineered crop- based agriculture
Plants, bacteria, fungi and animals whose genes have been altered by manipulation are called Genetically Modified
Organisms (GMO).

Advantages of Genetic Modification in plants.

o Made crops more tolerant to abiotic stresses (cold, drought, salt, heat)
o Reduce reliance on chemical pesticides (pest resistant crop)
o Helped to reduce post harvest losses.
o Increased efficiency of mineral usage by plants.
o Enhanced nutritional values of food e.g. vitamin A enriched rice.
Bt Cotton:

APPLICATIONS OF BIOTECHNOLOGY

PRODUCTION OF PEST RESISTANT PLANTS :-

(A) Bt Cotton :-
i) The soil bacterium Bacillus thuringiensis produces crystal proteins called Cry proteins that are toxic
to larvae of insects like tobacco budworm, armyworm, beetles and mosquitoes.
ii) The cry proteins exist as inactive protoxins and get converted into active toxin when ingested by the
insect, as the alkaline pH of the gut solubilises the crystals.
iii) The activated toxin binds to the surface of the epithelial cells of midgut and create pores.
iv) This causes swelling and lysis of cells leading to the death of the insect (larva).
v) The cry genes encoding this protein isolated from the bacterium and incorporated into several crop
plants like cotton , tomato,, corn, rice, soyabean etc.
vi) The proteins encoded by the following cry genes control the pest given against them :-
vii) Cry I Ac and Cry II Ab control cotton boll worms
viii) Cry I Ab controls corn borer.

88
Action of cry gene at a glance:

Bacteria Cry gene produces Crystal

Inactive in acidic pH
Bacillus thuringiensis protein (protoxin)

Isolation & insertion of

Eaten by insect Bt cotton
gene in cotton plant

Perforation in the
Gut of insect is alkaline Protoxin get activated
gut of insect

Hole is produced
Cotton plant protected Death of Insect in the gut of insect

(B) PROTECTION AGAINST NEMATODES

A nematode Meladogyne incognita infects tobacco plants and reduces their yield. This nematode is
controlled by a method called RNA interference :-
i) The specific genes ( in the form of cDNA ) from the parasite are introduced into the plant using
Agrobacterium as the vector.
ii) The genes are introduced in such a way that both sense/coding RNA and antisense RNA
(complementary to the sense / coding RNA) are produced.
iii) Since these two RNAs are complementary , they form a Double stranded RNA (dsRNA)
iv) This neutralises the specific RNA of the nematode by a process called RNA – interference.

89
 As a result the parasite cannot live in the transgenic host and the transgenic plant is
protected from the pest.

PEST RESISTANT PLANT - TOBACCO

HOST PLANT

INSERTION OF NEMATODE SPECIFIC GENE

USING Agrobacterium tumifaciens VECTOR

TRANSCRIPTION OF GENE TO MAKE

SENSE AND ANTI- SENSE m RNA

INFECTION OF HOST PLANT BY

NEMATODE- Meloidegyne incognitia

PREVENT TRANSLATION BY MAKING

OF ds RNA.- RNA Silencing

PROTECTION OF HOST

APPLICATIONS OF BIOTECHNOLOGY IN MEDICINE

1. The rDNA technology has been used in the production of safe and more effective therapeutic drugs.
2. The recombinant therapeutics do not induce unwanted immunological responses , that are
commonly observed with similar products isolated from non – human sources.
3. At present about thirty recombinant therapeutics have been approved for human use , of which
twelve are being marketed in India.

GENETICALLY ENGINEERED INSULIN ( HUMILIN )

a) Human insulin consists of two short polypeptide chains:- Chain A and Chain B , linked by
disulphide bridges.
b) Insulin is secreted as prohormone which has to be processed before it becomes a mature and
functional hormone .
c) The prohormone contains another polypeptide called C – peptide., which is removed during
maturation.
d) In 1983 , Eli Lilly , an American company , prepared two DNA sequences coding for chains A and
B of human insulin and introduced them into the E.coli to produce insulin.
90
e) The two chains produced were extracted and combined by creating disulphide bridges.

INSULIN PRODUCTION BY r-DNA TECHNOLOGY

Maturation of proinsulin to insulin

Maturation of Pro insulin to insulin:

https://www.youtube.com/embed/uST-u8flcow

HUMAN BODY

PROINSULIN – THREE POLYPEPTIDE CHAIN A,B,& C.

(Inactive )

MATURE INSULIN- ONLY CHAIN A & B

(C polypeptide is removed during processing)

ELI LILY (an US based company) SYNTHESISED CHAIN A & B

separately in E.coli USING rDNA TECHNOLOGY AND JOINED THEM
BY DISULFIDE BRIDGE.

https://www.youtube.com/watch?v=HSaFemVKtyc

91
 GENE THERAPY :
Gene Therapy is a collection of methods that allows correction of gene defect .
i) In this method , genes are inserted into the cells and tissues of an individual to correct certain
hereditary diseases.
ii) It involves the delivery of a normal gene into the individual or embryo to replace the defective
mutant allele of the gene.
iii) Viruses which attack the host and introduce their genetic material into the host are all used as
vectors.
iv) The first clinical gene therapy was given in 1990 to a four year old girl with adenosine deaminase
(ADA) deficiency.

ISOLATION AND CULTURE OF

LYMPHOCYTES

ISOLATION OF FUNCTIONAL
ADA GENE.

INSERTION OF ADA GENE USING c-DNA

IN LYMPHOCYTES.

INFUSION OF LYMPHOCYTES
IN THE BLOOD OF PATIENT

NEEDS PERIODIC INFUSION DUE TO

LIMITED LIFE SPAN OF WBC.

TREATMENT OF ADA DEFICIENCY

Traditional Method :- ADA deficiency can be cured by marrow transplantation in some children, but
it is not completely curable.
Modern Method :-
a) For gene therapy , lymphocytes were grown in a culture and functional ADA,c DNA is then
introduced into these lymphocytes. These lymphocytes are then transferred into the body of the patient .
The patient requires periodic infusion of such genetically engineered lymphocytes .
b) If a functional gene is introduced into the bone marrow cells at early embryonic stage, it would be a
permanent cure.
Molecular Diagnosis

92
 i) Recombinant DNA molecule and techniques like PCR are used for early diagnosis of disorders.

ii) Cloned genes when expressed to produce recombinant proteins , help in developing sensitive
diagnosis of techniques like ELISA.

iii) The cloned genes are also used as probes to detect the presence of complementary DNA strand.

A probe is a piece of a single stranded DNA that is tagged with a radioactive molecule andit
is used to find its complementary DNA by hybridisation. It is followed by detection of radioactivity by
autoradiography.
iv) Presence of normal or mutant gene can also be detected using such a method.

MOLECULAR DIAGNOSIS

MOLECULAR DIAGNOSIS

ELISA PCR

DETECT
ANTIGEN- ANTIBODY DETECT VERY DETECT HIV IN
MUTATION IN
ANTIBODY SYNTHESISED LOW AMOUNT OF SUSPECTED
CANCER
INTERACTION TO AGAINST THE DNA PATIENTS
PATIENTS.
DETECT HIV/ VIRAL PATHOGEN
INFESTATION

TRANSGENIC ANIMALS
Transgenic animals are those animals that have had their DNA manipulated to possess and express a
foreign gene.
Importance of Transgenic Animals :-
i) Transgenic animals can be specifically designed to allow the study of how genes are regulated and
how they affect the normal functions of the body and its developments. Eg. Information is obtained
about the biological role of insulin like growth factors.

Transgenic animals are designed to increase our understanding of how genes contribute to the
development of diseases. They are made to serve as models for human diseases.
iii) Transgenic animals that produce useful biological compounds can be created by introducing a
human protein used to treat emphysema.
eg. The first transgenic cow , Rosie ,produced the human protein enriched milk (2.4g/l).It also
contained human alpha lactalbumin, a more nutritionally balanced product for human babies.
93
 iv) Transgenic are being developed for use in testing the safety of vaccines (e.g. polio vaccine )
v) Transgenic animals with more sensitivity , to toxic substances are being developed to test the
toxicity of drugs.

BENEFITS OF TRANSGENIC ANIMALS:

BENEFITS OF TRANSGENIC ANIMALS

BIOLOGICAL STUDY OF VACCINE NORMAL PHYSIOLOGY CHEMICAL

PRODUCTS DISEASES SAFETY AND DEVELOPMENT SAFETY TESTING

Working of organ Transgenic

Alpha-1 Alpha- Cystic Fibrosis, Transgenic
Lactalbumin system is studied animals
antitryps cancer, Rheumatoid mice is used
In Rosie Arthritis.
are
(cow)

Substitute for
Emphysema mother’s milk
for infants.

STEM CELL TECHNOLOGY

* Definition :- Stem cells are the undifferentiated cells, which can divide (by mitosis) and
differentiate into specialised cells.
* Location :-
a) Embryonic stem cells are found in the inner cell mass of blastocyst and adult stem cells are found
in the bone marrow , blood , adipose tissue etc.
b) Stems cells are also obtained from umblical cord blood just after birth.
Properties of Stem Cells :-
a) Self – renewal or ability to multiply.
b) Potential to differentiate into specialised cells.
Example :- Bone marrow transplantation is an example of stem cell therapy.

ETHICAL ISSUES
* Genetic modification of organisms can have unpredictable / undesirable effects, when such
organisms are introduced into the ecosystem.
* The modification and use of such organism for public service has also resulted in problems with the
granting of patents.
GEAC (Genetic Engineering Approval Committee) :-
94
 The Indian Govt. has set up GEAC which is authorised:-
i) To make decisions regarding the validity of genetic modifications and
ii) The safety of introducing genetically modified organisms for public services.
BIOPIRACY
Biopiracy refers to the use of bioresources by multinational companies andother organisations
without proper authorisation form the countries and people concerned or without giving proper
compensation to the people / countries concerned.
The industrialised / developed nations are rich financially , but poor in biodiversity and traditional
knowledge, while the developing and underdeveloped countries are rich in bio resources and
traditional knowledge.Such developed countries often indulge in Biopiracy.
Example :- The genome of Basmati Rice , Neem and Haldi was illegally patented by US
Multinational companies illegally, however , these crops are of Indian Origin.

*******************************************************************************

95