BI 532: Experimental Principles in Molecular Biology

Thomas Jefferson University

April 14 & 18, 1997, "Yeast Transformation"

Charles Brenner, Ph.D.

charles.brenner@dartmouth.edu

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This week we will learn several recombinant DNA technologies that are routinely performed with laboratory strains of baker's yeast, Saccharomyces cerevisiae. You will learn the most basic facts about yeast chromosomal and extrachromosomal DNA, manipulation of which have made yeast transformation a powerful set of tools. In so doing, you will become conversant with the tricks of the trade and come to appreciate the "awesome power of yeast genetics." This page provides some of the background material for our discussion.

First, what are yeasts?

Second, how closely related are yeasts and humans?

Third, what is transformation? Transformation is getting an organism to take up foreign DNA without the use of viral sequences. The first report of transformation was by Avery, MacLeod and McCarty in 1944. The first report of transformation of yeast was from the laboratory of Gerry Fink. Protocols for yeast transformation have been refined since the early days and are available on the web.

We will discuss all the commonly used yeast shuttle vectors, including YIps, YRps, YCps, YEps and YACs. To appreciate the technology, you must learn what an ARS is, from its original description, and its midlife redefinition, to current understanding.

Yeast transformation vectors are used in a variety of interesting biological applications, including Mutant Allele Recovery, One-Step Gene Disruption, and One-Step Site-Directed Mutagenesis. Homework will require you to design an experiment to answer a biological question of interest to you that makes use of transformation of yeast cells.

Positions available.
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