Genetic Engineering 3

ISBN-13: 9781461570806 / Angielski / Miękka / 2012 / 173 str.

Robert Williamson

Opis

Like many genetic engineers, I have recently been receiving the atten tion of various venture capital companies, international drug houses and Members of Parliament. I will not discuss which of these approaches are most welcome, but it did cause me to consider the speed of advance in genetic engineering, and the implications of this rapid growth. There were few who anticipated it - only five years ago, most scientists thought applications would come at the end of the century, yet we see products such as insulin and interferon already available for clinical testing. In Europe in general and Britain in particular, this explosive growth in our own field has coincided with a general industrial depression and a marked reduction in funding for biomedical research. The brain drain from Britain is a serious matter, for we are losing the best of our younger scientists, on whom we would rely to train the next generation of molecular biologists. These volumes have come from British labs (mostly because I happen to be based in London, and my contacts and friends are here), and I feel that the quality of the con tributions also shows that our current research is of a high standard."

Spis treści

Plasmid and phage M13 cloning vectors.- I Introduction.- II Bacterial plasmids.- A Plasmid genes.- B Plasmid replication in E. coli.- C Transfer and mobilization.- III General purpose amplifiable vectors.- A Choice of vector.- B Choice of host cell.- C Selective markers.- D Regeneration of restriction sites.- E The pBR322 series.- IV Specialized vectors.- A Low copy number vectors.- B Vectors designed to detect transcription control signals.- C Direct selection vectors.- D Cosmids.- V Vectors designed to promote gene expression.- A Gene dosage.- B E. coli promoters.- C Fusion proteins.- VI Broad host range vectors.- A E. coli and Gram negatives.- B Bifunctional Bacillus-Escherichia vectors.- VII Single-stranded DNA phages as cloning vectors.- A Filamentous SS DNA phage biology.- B M13 vectors and their uses.- VIII Summary.- IX Acknowledgements.- X References.- Vectors based on bacteriophage lambda.- I Introduction.- II The lambda genome.- A The arrangement of the genes.- B The expression of lambda genes.- III Replication and maturation of lambda DNA.- A DNA structure.- B DNA packaging and size-selection.- C Maturation of recombinant phages.- D In vitro packaging of phage DNA.- IV The recognition of recombinant phages.- A Direct screens.- B Indirect screens.- C Positive selection for recombinant phages.- D Hybridization screening.- E Screens dependent on gene expression.- V The expression of genes cloned into lambda.- A Genes with their own promoter.- B Expression from PL.- C Expression from the late promoter.- D General considerations.- VI Making gene banks with lambda vectors.- VII Conclusions and future developments.- VIII Acknowledgements.- IX References.- Expression of cloned genes in eukaryotic cells using vector systems derived from viral replicons.- I Introduction.- A Why develop eukaryotic cloning systems?.- B What eukaryotic cloning systems are available?.- II Criteria for the design of animal virus vectors.- III Systems for the propagation of recombinant DNA molecules as virions.- A Simian virus 40.- B Human adenoviruses.- C Retroviruses.- IV Episomal vectors based on viral genomes.- A Introduction.- B Papillomavirus vectors.- C Episomal vectors based on Simian virus 40.- V Virus-based vectors carrying selectable genes.- A Introduction.- B The gpt system.- C The aminoglycoside phosphotransferase system.- D The dihydrofolate reductase system.- VI Vectors for the integration of exogenous DNA into chromosomal DNA.- VII Conclusions.- VIII Acknowledgements.- IX References.- A comprehensive list of cloned eukaryotic genes.- Key.- Genome clones.- cDNA clones.