RECOVERY OF VIRTUALLY FULL-LENGTH HIV-1 PROVIRUS OF DIVERSE SUBTYPES FROM PRIMARY VIRUS CULTURES USING THE POLYMERASE CHAIN-REACTION

In the course of the global pandemic, the human immunodeficiency virus type-1 (HIV-1) has established at least eight distinct genotypes in t he main (M), or prevalent, group of isolates, a variety of rare outlie r forms, and intergenotypic recombinants of group M viruses. This geno typic diversity has been documented, for the most part, by sequencing of subgenomic segments of the provirus. Using DNA from virus cultures on peripheral blood mononuclear cells (PBMC) and recent improvements o f the PCR technique, we have amplified virtually full-length HIV-I gen omes from genetic subtypes A through G of group M viruses and molecula rly cloned several of them. Resequencing of the complete genome of a p rototype strain after long PCR amplification and cloning has establish ed a PCR error rate of 0.14%. We also report the first complete PCR-de rived sequence of a U.S. clinical isolate of genotype a expanded only in primary PBMC; this provirus harbors a uniquely truncated V3 loop. ( C) 1995 Academic Press, Inc.