HUMAN-IMMUNODEFICIENCY-VIRUS REVERSE-TRANSCRIPTASE - ISOLATION AND SUBSTRATE-SPECIFICITY

Citation
Ta. Rozovskaya et al., HUMAN-IMMUNODEFICIENCY-VIRUS REVERSE-TRANSCRIPTASE - ISOLATION AND SUBSTRATE-SPECIFICITY, Molecular biology, 27(3), 1993, pp. 376-383
Citations number
27
Categorie Soggetti
Biology
Journal title
ISSN journal
00268933
Volume
27
Issue
3
Year of publication
1993
Part
2
Pages
376 - 383
Database
ISI
SICI code
0026-8933(1993)27:3<376:HR-IAS>2.0.ZU;2-X
Abstract
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.