Papers by Geoffrey Turner
Bio/technology (Nature Publishing Company), 1990
A cosmid clone containing the putative penicillin biosynthetic gene cluster from Penicillium chry... more A cosmid clone containing the putative penicillin biosynthetic gene cluster from Penicillium chrysogenum was used to transform the related filamentous fungi Neurospora crassa and Aspergillus niger, which do not produce beta-lactam antibiotics. Both of the transformed hosts contained intact P. chrysogenum DNA derived from the cosmid clone and produced authentic penicillin V. Assays of penicillin biosynthetic enzyme activity additionally demonstrated that they possessed delta-(L-alpha-amino-adipyl)-L-cysteinyl-D-valine synthetase (ACVS), isopenicillin N synthetase (IPNS) and acyl coenzyme A:6-aminopenicillanic acid acyltransferase (ACT) activity. The data suggests that genes encoding all the enzymes necessary for the biosynthesis of penicillin from amino acid precursors are closely linked in P. chrysogenum and constitute a gene cluster.
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Current genetics, 2003
Non-ribosomal peptide synthetases, polyketides and fatty acid synthetases have a modular organisa... more Non-ribosomal peptide synthetases, polyketides and fatty acid synthetases have a modular organisation of multi-enzymatic activities. In all of them, the acyl or peptidyl carrier proteins have 4'-phosphopantetheine (P-pant) as an essential prosthetic group. This is added by 4'-phosphopantetheinyl transferases (PPTases) that derive the P-pant group from coenzyme A. While many PPTases of varying specificity have now been isolated from a number of bacteria, a filamentous fungal PPTase has yet to be characterised. Through database searching of the Aspergillus fumigatus genome sequence against Sfp from Bacillus subtilis, we identified a unique sequence which appears to encode a PPTase, as deduced from conserved residues considered important in PPTases. The PPTase candidate was used to search the NCBI data base and an unexpected homologue in A. nidulans was identified as npgA. Mutations in this gene (cfwA/ npgA) were identified previously as leading to defects in growth and pigment...
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Revista Iberoamericana de Micología, 2005
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PLoS Genetics, 2008
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Physiotherapy, 2001
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Nature, 2005
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MGG Molecular & General Genetics, 1975
Three extranuclear mitochondrial mutations in Aspergillus nidulans, (oliA1), (camA1) and (cs67), ... more Three extranuclear mitochondrial mutations in Aspergillus nidulans, (oliA1), (camA1) and (cs67), were used as markers in sexual crosses to provide information on the frequencies of transmission and recombination of the mitochondrial genome. Any individual perithecium contained ascospores of only one extranuclear genotype. Using mono-, bi- and trifactorial crosses it was found that all three markers could be recovered from the progeny, although the transmission frequencies were different for each marker. This bias was present irrespective of the nuclear background or the presence of selective agents in the medium on which the cross was established. These findings enable a series of "transmission strength" to be established, as shown below:-- (camA1) greater than (cs67, camA1) greater than (+) = (cs67) greater than (oliA1, cs67) greater than (oliA1) greater than (oliA1, camA1). However, the numbers of recombinants isolated were so variable as to make this form of analysis unsuitable for mapping the mitochondrial genome.
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MGG Molecular & General Genetics, 1989
The isopenicillin N synthetase (IPNS) gene has been isolated from wild-type Penicillium chrysogen... more The isopenicillin N synthetase (IPNS) gene has been isolated from wild-type Penicillium chrysogenum and used as a probe to detect the equivalent gene on Southern blots of genomic DNA from a mutant producing high levels of penicillin. The IPNS gene in this strain is contained within a region of DNA of wild-type restriction pattern that extends for at least 39 kb and is present at between 8 and 16 copies. The steady state level of IPNS mRNA in the mutant producing high levels of penicillin is between 32- and 64-fold of that of the wild type, suggesting that the rate of transcription of some or all of the copies has been increased. In addition we have also shown that both the IPNS mRNA and enzyme is present throughout the growth phase in both strains under the culture conditions used. IPNS enzyme activity is greatly increased in the strain with the high penicillin titre.
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Manual Therapy, 2000
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Fungal Genetics and Biology, 2009
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Fungal Genetics and Biology, 2004
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Fungal Genetics and Biology, 2011
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Current Genetics, 1988
An amplified sequence of mitochondrial DNA from a ragged (rgd) mutant of Aspergillus amstelodami ... more An amplified sequence of mitochondrial DNA from a ragged (rgd) mutant of Aspergillus amstelodami has been shown to exist in multimeric circular form, suggesting that it is excised from the genome and can exist independently of it. This sequence has replicative (ARS) activity in Saccharomyces cerevisiae, and a subfragment responsible for this activity has been identified and sequenced. A homologous sequence from Aspergillus nidulans mtDNA also has ARS activity in S. cerevisiae. Both A. amstelodami and A. nidulans ARS elements have been incorporated into the integrative transformation vector pDJB1 and the derived vectors used to transform A. nidulans. Inclusion of the A. nidulans ARS element enhanced the transformation frequency 5-fold relative to pDJB1. No increase in transformation frequency was evident with the ARS element from A. amstelodami. The stability of transformants was variable but in comparison to pDJB1, ARS-containing plasmids were mitotically unstable in A. nidulans. Although plasmid DNAs could be rescued in Escherichia coli from undigested transformant DNA, no freely replicating plasmids were detected by Southern hybridisation.
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Current Genetics, 1987
The Aspergillus nidulans acetamidase gene (amdS) has been used to transform Penicillium chrysogen... more The Aspergillus nidulans acetamidase gene (amdS) has been used to transform Penicillium chrysogenum at low frequency. Several transformants were tested and shown to be mitotically stable. Southern blot analysis indicated that transforming DNA had integrated into the chromosomal DNA, possibly at multiple sites.
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Current Genetics, 1988
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Current Biology, 2001
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Current Biology, 2001
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Current Biology, 1999
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Applied and Environmental Microbiology, 2003
Here we report the identification of the -lactam biosynthesis genes pcbAB and pcbC from a cosmid ... more Here we report the identification of the -lactam biosynthesis genes pcbAB and pcbC from a cosmid genomic DNA library of the marine fungus Kallichroma tethys. A BLAST homology search showed that they share high sequence identity with the -(L--aminoadipyl)-L-cysteinyl-D-valine (ACV) synthetases and isopenicillin N synthases, respectively, of various fungal and bacterial -lactam producers, while phylogenetic analysis indicated a close relationship
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Trends in Biotechnology, 1998
Penicillin production by Penicillium chrysogenum is not only commercially important but arguably ... more Penicillin production by Penicillium chrysogenum is not only commercially important but arguably the most intensively investigated secondary-metabolic pathway in fungi. Isolation of the structural genes encoding the three main penicillin-biosynthetic enzymes has stimulated the use of molecular approaches to optimize yield and permitted genetic analysis of current production strains, which are themselves the products of 50 years of strain and process improvement. Parallel studies on the penicillin-producing genetic model organism Aspergillus nidulans are now addressing questions about the genetic regulation of primary and secondary metabolism, the compartmentalization of biosynthesis and the excretion of the end products.
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Papers by Geoffrey Turner