ENHANCED PRODUCTION OF HIV-1 PROTEASE IN E COLI

HUMAN IMMUNODEFICIENCY VIRUS TYPE I (HIV-1) IS THE ETIOLOGICAGENT OF ACQUIRED IMMUNE DEFICIENCY SYNDROME (AIDS). THIS PROJECT'S LONG-TERM OBJECTIVE IS TO STUDY THE ACTION OF THE HIV-1 PROTEASE TO DEVELOP SPECIFIC PROTEASE INHIBITORS THAT MIGHT BE ABLE TO HALT THE VIRAL REPLICATION. THIS STUDY IS POSSIBLE ONLY WITH THE AVAILABILITY OF LARGE QUANTITIES OF BIOLOGICALLY ACTIVE HIV-1 PROTEASE. PHASE I EFFORTS WILL DEVELOP A RECOMBINANT DNA SYSTEM TO PRODUCE THE PROTEASE. THE APPROACH TO BE USED WILL INVOLVE THE ASSEMBLY OF A SYNTHETIC GENE CODING FOR THE HIV-1 PROTEASE. THE GENE WILL BE CLONED INTO A PLASMID AND EXPRESSED IN ESCHERICHIA COLI. THE PRODUCTION LEVEL OF THE GENE PRODUCT WILL BE OPTIMIZED SO THAT THE PROTEASE CAN BE ECONOMICALLY PRODUCED. HIGH-LEVEL EXPRESSION OF THE PROTEASE WILL ENABLE PHASE II RESEARCH TO FOCUS ON THE E. COLI FERMENTATION SCALE-UP, PURIFICATION, AND RENATURATION OF THE PROTEASE TO YIELD BIOLOGICALLY ACTIVE HIV-1 PROTEASE.