Development of a yeast-based recombination cloning/system for the analysis of gene products from diverse human...

A recent shift from studies on a few subtype B laboratory human immunodeficiency virus type 1 (HIV-1) clones to analyses of extremely diverse primary HIV-1 isolates from different subtype requires the development of a rapid and generic cloning technique. This report describes the use of gap repair/recombination in yeast to shuttle env, gag, and pol genes from diverse HIV-1 subtypes into a DNA vector that can be amplified in bacteria and can express the gene of interest in mammalian cells. These diverse HIV-1 genes have also been introduced into an infectious clone to produce chimeric viruses that are useful for studies on drug susceptibility, receptor binding and fitness.