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Principles and Practice of Recombinant DNA Research with Yeast

David Botstein, Ronald W. Davis


Recombinant DNA research shows great promise in furthering understanding of yeast biology by making possible the analysis and manipulation of yeast genes not only in the test tube but also in yeast cells. The intent of this paper is to summarize the major features of current technology and to outline some implications for the immediate future of yeast molecular biology. There now exist simple and general methods for isolating and amplifying virtually any yeast gene, although these methods generally require an intermediate step in Escherichia coli. Powerful and exquisitely sensitive hybridization methods have been developed that allow direct physical analysis of any chromosomal region containing a gene that has been molecularly cloned. Most importantly, it is now possible to return to yeast, by transformation with DNA, cloned genes using a variety of selectable marker systems developed for this purpose. These technological advances have combined to make feasible truly molecular as well as classical genetic manipulation and analysis in yeast.

The many uses to which these new tools have already been put have recently been reviewed by Olson (1981), and the results that have been obtained are to be found scattered in many of the papers in this volume. The biological problems that have been most effectively addressed by recombinant DNA technology are ones that have the structure and organization of individual genes as their central issue. Thus, the reader will find elsewhere in this volume that many signal advances in the understanding of such diverse subjects as the switching...

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