A yeast expression vector and leucine selection in Escherichia coli to aidin the identification of novel genes

The complete sequencing of the Saccharomyces cerevisiae genome provides a p owerful toot for studying and elucidating essential cellular processes. To aid in the application of this resource to investigations into the molecula r mechanisms of endoplasmic reticulum-associated protein degradation, a sim ple procedure was designed to generate a unique 2-mum LEU2-selectable yeast expression vector. Putative genes easily inserted into this vector are und er control of the ADH1 promoter and transcription terminator sequences. Fur thermore, a LEU2 selection in both yeast and Escherichia coli was used to a llow the isolation of underrepresented plasmid from a pool of multiple vect ors. Together, these advances in technology will be useful in the systemati c analysis of novel yeast gene function, (C) 2001 Academic Press.