A yeast expression vector and leucine selection in Escherichia coli to aid in the identification of novel genes.
The complete sequencing of the Saccharomyces cerevisiae genome provides a powerful tool for studying and elucidating essential cellular processes. To aid in the application of this resource to investigations into the molecular mechanisms of endoplasmic reticulum-associated protein degradation, a simple procedure was designed to generate a unique 2-microm LEU2-selectable yeast expression vector. Putative genes easily inserted into this vector are under control of the ADH1 promoter and transcription terminator sequences. Furthermore, a LEU2 selection in both yeast and Escherichia coli was used to allow the isolation of underrepresented plasmid from a pool of multiple vectors. Together, these advances in technology will be useful in the systematic analysis of novel yeast gene function.
Palmer
E A
EA
Department of Biology, University of Nevada, Reno, 89557, USA.
Kruse
K B
KB
McCracken
A A
AA
D050539:3-Isopropylmalate Dehydrogenase; D000426:Alcohol Dehydrogenase / Q000235:genetics; D000429:Alcohol Oxidoreductases / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D005798:Genes, Bacterial; D005822:Genetic Vectors; D007930:Leucine / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000235:genetics*
D000429:Alcohol Oxidoreductases; D000426:Alcohol Dehydrogenase; D050539:3-Isopropylmalate Dehydrogenase; D007930:Leucine