Methods for the production, by genetic manipulation, of a recombinant enzymatically active HIV-specific protease and the precursor molecules of the group-specific antigens (gag) and of the polymerase (pol) and use of these products in mixtures for testing HIV protease-specific inhibitors

摘要

This invention comprises the simple and non-hazardous possibility of testing the activity of specific protease inhibitors, both in vitro and in vivo, with the aid of enzyme and substrate produced by genetic manipulation. This invention describes the production, by genetic manipulation, of polypeptide precursor molecules (gag precursor molecule and pol precursor molecule) as well as the expression of an enzymatically active HIV protease. The use of the said proteins in both in vitro mixtures (as protein extracts) and in vivo mixtures (expression in prokaryotic or eukaryotic cells) permits the effect of specific inhibitors on HIV protease to be determined in simple test mixtures. At the same time, the test mixtures described can be used to check the penetration of such substances into cells. The described test mixtures use exclusively HIV-authentic protein sequences. This makes it possible to avoid the elaborate and not entirely non-hazardous purification of corresponding proteins from virus-infected cell cultures or isolated virus particles.