Screening apoptosis inhibitors using the BAX gene

摘要

A method of screening cDNA encoding putative apoptosis inhibitors in yeast cells expressing the pro-apoptotic BAX gene is described. Saccharomyces cerevisiae are transformed with vectors containing the BAX gene and a promoter, and candidate cDNA's. BAX gene expression is induced by exposing the cells to a substance which activates the promoter. cDNA is recovered from those cells which do not display apoptosis and introduced into a mammalian cell, for example COS cells, which are then challenged with a proapoptotic protein. Those cells which do not exhibit apoptosis confirm the inhibitory capacity of the cDNA. The restoration of cell growth in yeast expressing BAX genes by Bcl-2 and Bcl-xL is described. The BAX-delta gene lacks the region coding for a C-terminal membrane anchor. Bax-delta expressing cells can be recovered by transfection with a human cerebellum cDNA library but not by Bcl-2 or Bcl-xL.