Eukaryotic expression of enzymatically active human immunodeficiency virustype 1 reverse transcriptase

Reverse transcriptase of human immunodeficiency virus type 1 is a vitalenzy me in the HIV-1 replication cycle and an attractive target of attempts to a rrest a primary viral infection. We designed a vector for eukaryotic expres sion of the 66 kDa subunit of reverse transcriptase under the control of th e immediate early cytomegalovirus promoter, Efficient transient expression of the 66 kDa subunit of reverse transcriptase was achieved in a variety of cells. Immunostaining of the transfected cells revealed the cytoplasmatic localization of reverse transcriptase, Reverse transcriptase activity was d etected in all transfected cell lines. Injection of this plasmid encoding t he 66 kDa subunit of reverse transcriptase into mice resulted in strong rev erse transcriptase-specific immune responses indicating that the 66 kDa sub unit of reverse transcriptase is expressed in vivo. Sera from DNA-immunized mice inhibited reverse transcription in vitro. (C) 1999 Federation of Euro pean Biochemical Societies.