WILD-TYPE AND MUTANT HIV-1 AND HIV-2 TAT PROTEINS EXPRESSED IN ESCHERICHIA-COLI AS FUSIONS WITH GLUTATHIONE-S-TRANSFERASE

Human immunodeficiency virus type 1 (HIV-1) and HIV-2 encode related t ranscriptional activators known as Tat-1 and Tat-2, respectively, that are required for efficient viral replication. The Tat proteins have b een studied extensively, and it appears that their mechanism of action is unique to the primate immunodeficiency viruses or a few distantly related lentiviruses. Here we describe a collection of 24 wild-type an d mutant Tat-1 and Tat-2 proteins that are expressed in Escherichia co li as fusions with glutathione S-transferase (GST). The GST-Tat fusion s can be used for biochemical studies after simple purification from E . coli lysates in a single step under nondenaturing conditions. The av ailability of these GST-Tat fusions should be useful to investigators examining biochemical properties of Tat-1 and Tat-2 proteins. E. coli cultures harboring GST-Tat fusions described here are available throug h the National Institute of Health AIDS Research and Reference Reagent Program.