Yeast Selection of Drug Resistant HIV Protease Variants

The therapeutic effects of drugs against HIV are overcome by drug resistant HIV variants. We descridrug resistance 'trap' for discovery by rapid positive selection of drug resistant HIV protease variadaptation of the yeast interaction trap/two hybrid system (Brent and Ptashne, Cell 43: 729; Fields340:245). Transcription of specific reporter genes or selectable marker genes in yeast are to be putelement transcription activator protein. Genetic engineering will be used to construct a hybrid actitwo activator elements are joined by a segment of HIV pol polyprotein. The HIV portion of the hybridprotease and one or more of its cleavage substrate sites. Activation of transcription by this hybriddependent upon the activity of the protease. Media are described which will allow growth of the drugcells only if the protease is active or only if the protease is inactive. Large libraries of mutantscreened or selected from to discover HIV protease variants which are active despite the presence ofdrugs.