Cloning Regulated Yeast Genes from a Pool of lacZ Fusions

Publisher Summary This chapter discusses cloning regulated yeast genes from a pool of lacZ fusions. Gene fusions have been used during the past decade to study such diverse problems as transcriptional regulation, translational regulation, and protein localization. In prokaryotes, various genetic methods have been employed to create operon and protein fusions. The active, regulated gene fusions constructed by the procedures described below contain a yeast DNA fragment with sequences for the control and initiation of transcription and translation joined to a lacZ fragment from which the 5’ portion including the first few N-terminal amino acid codons of the gene have been deleted. A library of random yeast gene:lacZ fusions in yeast transformants was constructed and screened for regulated gene fusions.