Abstract
A system is described that facilitates the selection of yeast promoters of unknown origin using a functional heterologous gene assay. A multicopy plasmid was constructed with a unique cloning site 5′ to the Herpes Simplex type 1 thymidine kinase gene (TK). A yeast genomic DNA library was introduced into this site and used to transform yeast (which naturally lacks a TK gene). Only transformants carrying DNA fragments with promoter activity can express TK activity and thereby grow on medium containing thymidine and folate antagonists. On the basis of TK assays of yeast cell homogenates from some 100 TK + transformants, a particularly promising plasmid containing a yeast promoter was selected for further analysis.
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