Generation and characterization of the human immunodeficiency virus type 1 mutants

Abstract

Mutations were introduced by recombinant DNA techniques into 9 genes of an infectious molecular clone of human immunodeficiency virus type 1. The 24 mutants generated were characterized biochemically and biologically by transfection and infection experiments. None of the mutants which have mutations ingag (p17, p24, and p15 regions),pol (protease, reverse transcriptase, and endonuclease domains),env (gp120 region),tat, orrev were infectious, whereasvif, vpr, vpu, some ofenv (gp41) andnef mutants could grow in human CD4⁺ cells to various degrees. Of the non-infectious mutants, only endonuclease (pol) and gp41 mutants exhibited normal phenotypes with respect to the production of functional reverse transcriptase, the expression ofgag, pol, andenv proteins, and the generation of progeny virions, when examined in transient assays. All infectious mutants killed the CD4⁺ cells with the exception of a mutant carrying a defect in thevif gene.