Module 1

Definition

Genetic engineering (genetic modification/manipulation) is the direct modification of an

organism’s DNA to alter its characteristics.

Key Concepts

 DNA Code: All life forms share the same DNA code, allowing genes from one organism
to function in another.
 Plasmids: Circular DNA in bacteria commonly used in genetic engineering.
 Recombinant DNA (rDNA): Artificially created DNA by combining genes from
different organisms.

Genetic Engineering Process

1. Selection of Gene – Identify a gene of interest.
2. DNA Extraction – Isolate DNA from the organism.
3. Gene Cutting – Use restriction enzymes ("molecular scissors") to cut the DNA.
4. Gene Insertion – Insert the desired gene into a plasmid.
5. Transformation – Introduce recombinant DNA into a host organism (e.g., bacteria).
6. Replication & Expression – The host organism multiplies and expresses the gene.

Example: Insulin Production Using Genetic Engineering

1. Extract plasmid from bacteria/yeast.
2. Cut plasmid using restriction enzymes.
3. Insert the human insulin gene.
4. Insert the modified plasmid into bacteria/yeast.
5. Bacteria/yeast reproduce, producing insulin.
6. Insulin is harvested, purified, and packaged.

Recombinant DNA Technology

 Developed by Boyer & Cohen (1973).
 Uses vectors (plasmids/viruses) to introduce genes into organisms.
 Key enzymes:
 o Restriction enzymes – Cut DNA at specific sites.
o Ligase enzyme – Joins DNA fragments together.
 Gene Transfer Methods:
o Physical: Electroporation, microinjection.
o Chemical: Calcium chloride, PEG-mediated.
o Biological: Virus-mediated (transduction).

Applications of Genetic Engineering

Medicine & Health

 Vaccines – Development of safer, more effective vaccines.

 Insulin Production – Used for diabetes treatment.
 Human Growth Hormone (HGH) – Treats growth disorders.
 Interferons – Used in cancer treatment.
 Antibiotics – Mass production of drugs like penicillin.
 Monoclonal Antibodies – Used in diagnostics and cancer therapy.
 Gene Therapy – Alters defective genes to treat diseases.
 DNA Fingerprinting – Used for forensic identification.
 PCR (Polymerase Chain Reaction) – Amplifies DNA for analysis.

Agriculture

 Herbicide Tolerance – Crops resistant to herbicides (e.g., glyphosate-resistant

soybeans).
 Insect Resistance – Bt crops (e.g., Bt cotton, Bt corn) produce natural insecticides.
 Stress Tolerance – Crops engineered to withstand drought, salinity, and extreme
temperatures.
 Disease Resistance – Plants modified to resist viruses and bacteria.
 Improved Nutrition – Golden rice enriched with Vitamin A.

Environment

 Bioremediation – Bacteria engineered to clean oil spills and degrade toxins.

 Nitrogen Fixation – Engineered microbes improve soil fertility.
 Waste Treatment – Breakdown of pollutants using modified bacteria.

Energy

 Biofuels – Genetically modified algae for biodiesel production.

 Ethanol Production – Yeast engineered for efficient alcohol fermentation.

Food Industry
  Dairy Products – Enzymes like chymosin (for cheese production).

Key Historical Milestones

 1966 – Nirenberg & Matthaei break the genetic code.
 1970 – First restriction enzyme discovered.
 1972 – Paul Berg creates the first recombinant DNA molecule.
 1973 – Boyer & Cohen develop recombinant DNA technology.
 1983 – Kary Mullis invents PCR.
 2000 – Gateway Cloning System developed for efficient DNA transfer.

Ethical Considerations

 Genetic Modification Risks – Potential unintended mutations.

 GMO Safety – Debate over human and environmental impact.
 Gene Therapy Ethics – Issues surrounding genetic modifications in humans.

Module 2

I. The Origin of Life

A. Early Experiments and Theories

 Louis Pasteur’s Experiments (1864):

o Demonstrated that microbes come from preexisting life (biogenesis).
o Debunked the idea of spontaneous generation—organisms appearing
miraculously from non-living matter.
 The Riddle:
o If life can only arise from life, how did the first living organisms appear on Earth?

B. Theories for Life’s Beginnings

 Organic Molecules & Complexity:

 o Life required the formation of organic molecules (containing carbon).
o These molecules eventually formed more complex compounds such as proteins,
carbohydrates, and nucleic acids.
 Oparin-Haldane Hypothesis:
o Proposed that lightning, the sun’s heat, and Earth’s internal heat triggered
chemical reactions.
o Resulted in a “primitive soup” in the early oceans, rich in organic molecules.
 Miller-Urey Experiment (1953):
o Simulated early Earth conditions by mixing water vapor, ammonia, methane, and
hydrogen with electricity.
o Produced organic molecules (e.g., amino acids and sugars) supporting the Oparin-
Haldane hypothesis.
 Alternative Hypothesis: Hydrothermal Vents:
o Deep-sea vents (hot and alkaline conditions) might have provided the right
environment for organic compounds to form.
 Formation of Protocells:
o Research (e.g., Sidney Fox’s work) showed that under certain conditions,
complex molecules can form protocells—organized structures that grow and
divide.

II. The Fossil Record & Geologic Time

A. Fossils and What They Tell Us

 Definition:
o Fossils are the preserved remains or traces of organisms from the past.
 Importance:
o Show evidence of past life, including behavior, size, diet, and environmental
conditions.
o Indicate that about 95% of all species that ever lived are now extinct.
 Formation:
o Commonly found in sedimentary rocks (formed by compression of mud, sand, or
clay).
o Also discovered in ice and amber.

B. Radiometric Dating

 Method:
o Measures the decay rate of radioactive isotopes (e.g., potassium-40, carbon-14).
o Uses half-lives as a “clock” to determine the age of rocks and nearby sedimentary
layers.

C. The Geologic Time Scale

  Four Major Eras:
1. Precambrian:
 Covers ~87% of Earth’s history.
 Dominated by one-celled organisms; multicellular life appeared late.
2. Paleozoic:
 Significant increase in organism diversity.
 Features the Cambrian explosion and the emergence of fishes, amphibians,
reptiles.
 Ends with a mass extinction (loss of ~90% of ocean species and ~70% of
land species).
3. Mesozoic:
 Divided into Triassic, Jurassic, and Cretaceous periods.
 Rise of dinosaurs, early mammals, and the evolution of birds from
dinosaurs.
 Ended with the extinction of the dinosaurs (possible meteor impact).
4. Cenozoic:
 Began about 65 million years ago.
 Known as the Age of Mammals, culminating in the evolution of modern
humans (appeared ~200,000 years ago).

III. Darwin’s Contributions and Evolutionary Theory

A. Darwin’s Journey and Observations

 Voyage of the Beagle:

o Observations (e.g., Galápagos finches) led Darwin to consider that species adapt
over time.
 Darwin’s Theory (Published in 1859):

1. Common Descent:
 All species are related and descended from common ancestors.
2. Natural Selection:
 The mechanism by which evolution occurs.

B. Natural Selection Explained

 Key Points:
o Works on existing genetic variation within populations.
o Favors individuals best suited for their environment (those that are more likely to
survive and reproduce).
o Does not produce perfect organisms but rather those “good enough” for current
conditions.
 Examples in Nature:
 o Galapagos Finches:
 Variation in beak size correlates with food availability; environmental
changes drive evolution in beak morphology.
o Peppered Moths:
 In polluted areas, darker moths are favored (camouflage on soot-darkened
trees), while in unpolluted areas, lighter moths dominate.
 Artificial Selection:
o Human-driven breeding of crops and animals shows rapid evolutionary changes,
supporting the power of natural selection.

IV. Genetic Variation and the Mechanisms of Evolution

A. Sources of Genetic Variation

 Mutations:
o Random changes in DNA that create new alleles.
 Other Sources:
o Gene duplication.
o Crossing over and independent assortment during sexual reproduction.

B. The Hardy-Weinberg Principle

 Concept:
o Predicts that allele and genotype frequencies remain constant (equilibrium) in a
large, randomly mating population with no mutation, gene flow, or natural
selection.
 Mathematical Representation:
o For two alleles, p and q:
 p² = frequency of one homozygote.
 2pq = frequency of heterozygotes.
 q² = frequency of the other homozygote.
 Deviations:
o Any deviation from these predicted frequencies indicates that evolutionary
processes are at work.

C. The Five Agents of Evolutionary Change

1. Mutation:
o Introduces new genetic variants.
2. Gene Flow:
o Movement of alleles between populations (migration).
3. Nonrandom Mating:
 o When individuals select mates based on specific traits (assortative or
disassortative mating).
4. Genetic Drift:
o Random changes in allele frequencies, especially in small populations.
5. Natural Selection:
o Differential reproductive success of individuals based on their traits.

V. Types of Natural Selection

A. Stabilizing Selection

 Definition:
o Favors average individuals, reducing variation.
 Example:
o In a spider population, average-sized individuals may be less noticeable to
predators and better at finding food.

B. Directional Selection

 Definition:
o Favors one extreme of the trait distribution.
 Example:
o Woodpeckers with longer beaks may have an advantage if their food source is
located deeper in trees.

C. Disruptive Selection

 Definition:
o Favors both extremes, potentially leading to two distinct phenotypes.
 Example:
o Limpets with very light or very dark shells may be favored depending on the rock
color, while those with intermediate colors are more visible to predators.

Summary

 Origins of Life:
o Life began from preexisting organic molecules that became increasingly complex
through chemical reactions on early Earth.
 Fossils & Geologic Time:
o Fossils record Earth’s biological history, with radiometric dating providing a
timeline of major eras and events.
  Darwin and Evolution:
o Darwin’s observations led to the theory of evolution by natural selection, which is
supported by modern evidence from finches, moths, and artificial selection.
 Genetic Variation & Mechanisms:
o Genetic variation, essential for evolution, arises through mutations and other
genetic processes. The Hardy-Weinberg principle helps detect when evolution is
occurring.
 Agents of Evolution:
o Evolution is driven by mutation, gene flow, nonrandom mating, genetic drift, and
natural selection, each contributing to changes in allele frequencies over time.

Module 3