The isolation of human immunodeficiency virus (HIV) reverse transcript
ase produced in bacteria [E. coli RRi strain (pRC-RT, pRK248cIts)] is
described. Substrate properties of 2'-deoxyribonucleoside 5'-triphosph
ate analogs studied previously in cell-free systems with avian myelobl
astosis virus and Moloney murine leukemia virus reverse transcriptases
were examined in vitro with HIV reverse transcriptase. The substrate
properties of new 2'-deoxyadenosine 5'-triphosphate analogs-2',3'-dide
oxy-2',3'-didehydro- and 2',3'-dideoxytubercidin 5'-triphosphates-were
examined. The relative efficiency of incorporation of different 2'-de
oxyribonucleoside 5'-triphosphate analogs into the DNA chain was evalu
ated. It was shown that HIV reverse transcriptase cloned in E. call an
d purified by the described method exhibits selectivity toward various
2'-deoxyribonucleoside 5'-triphosphates that is characteristic of the
previously studied reverse transcriptases including the native viral
one. This property permits the employment of the cloned enzyme in diff
erent model systems.