A comprehensive collection of readily reproducible techniques for the manipulation of recombinant plasmids using the bacterial host E. coli. The authors describe proven methods for cloning DNA into plasmid vectors, transforming plasmids into E. coli, and analyzing recombinant clones. They also include protocols for the construction and screening of libraries, as well as specific techniques for specialized cloning vehicles, such as cosmids, bacterial artificial chromosomes, l vectors, and phagemids. Common downstream applications such as mutagenesis of plasmids, recombinant protein expression, and the use of reporter genes, are also described.

This concise reference focuses on the manipulation of plasmids in Escherichia coli.

This concise reference focuses on the manipulation of plasmids in Escherichia coli.

This book was originallyconceived in the form ofa second edition ofa volume published in 1980 in Chapman and Hall's 'OutllneStudies in Biology' series and entitled Genetic Engineering - Cloning DNA. It very rapidly became apparent that with the impact ofrecombinant DNA techniques being feIt in so many areas ofblology, it was going to be difficultifnotimpossible to keepthe bookwithin the space confines of these little monographs. The stays were therefore loosened and the book expanded comfortably to its present size. I hope that this extra space has allowed me to clarify sections ofthe text that were 'heavy going' in the earlierversion. Theextraspace has certainlyallowed me to cover topics that were not mentioned at all in the earlier book. These are primarily to be found in Chapters 7 and 8, which cover the rapid advances that have been recently made in the use ofplantand animal cells as hosts for recombinant DNAmolecules. The develop ment ofother vectors has certainly not stood still over the past four years. This has necessitated a thorough revision ofChapters 3 and 4, which deal with bacteriophage and bacterial plasmid vectors. Numerous techniques for in vitromutagenesis have now been tried and tested allowing me to givecomprehensive coverage ofthisarea in Chapter 2 along with the biochemical techniques used to construct recombinant DNA molecules. Readers with some background knowledge of the approaches to gene cloning will be able to go straight toapart ofthe book in whichthey are specificallyinterested.

A comprehensive collection of readily reproducible techniques for the manipulation of recombinant plasmids using the bacterial host E. coli. The authors describe proven methods for cloning DNA into plasmid vectors, transforming plasmids into E. coli, and analyzing recombinant clones. They also include protocols for the construction and screening of libraries, as well as specific techniques for specialized cloning vehicles, such as cosmids, bacterial artificial chromosomes, l vectors, and phagemids. Common downstream applications such as mutagenesis of plasmids, recombinant protein expression, and the use of reporter genes, are also described.

This book resulted from presentations at an international conference on bacterial p1asmids held January 5-9, 1981 in Santo Domingo, Dominican Republic. This was the first meeting of its kind in the Southern Hemisphere. The meeting place was selected for its relaxed and comfortable climate, conducive to interactions among participants. More importantly the locale facilitated the participation of nearby Latin American clinical and research scientists who deal directly with the health manifestations of pathogenic p1asmids. Diseases and socio-economic practices of developing countries exist in the Dominican Republic whose scientific community could directly benefit from having the meeting there. The book includes the talks as well as extended abstracts of poster presentations from the meeting. This combination, which provides readers with reviews as well as recent findings, captures the full scientific exchange which took place during the 5-day meeting. As one indication of pathogenicity related to p1asmids, the conferees were surveyed for gastro-intestina1 problems during and after their stay in the Dominican Republic. The results are summarized at the end of this book.