Characterization of recombinant integrase of human immunodeficiency virus type 1 (isolate Bru)

被引：2
|
作者：
Semenova, EA [1 ]
Gashnikova, NM [1 ]
Il'ina, TV [1 ]
Pronyaeva, TR [1 ]
Pokrovsky, AG [1 ]
机构：
[1] State Res Ctr Virol & Biotechnol Vector, Inst Mol Biol, Koltsov 630559, Novosibirsk Reg, Russia
来源：
BIOCHEMISTRY-MOSCOW | 2003年 / 68卷 / 09期
基金：
俄罗斯基础研究基金会;
关键词：
integrase; human immunodeficiency virus type 1; isolate Bru; aurintricarbonic acid;
D O I：
10.1023/A:1026060512380
中图分类号：
Q5 [生物化学]; Q7 [分子生物学];
学科分类号：
071010 ; 081704 ;
摘要：
Integration of the human immunodeficiency virus type 1 (HIV-1) DNA into the human genome requires the virus-encoded integrase protein. The recombinant integrase protein of HW-1 (isolate Bru) was prepared by constructing a plasmid based on pET-15b encoding the integrase gene. Integrase of HfV-1 was purified using a bacterial expression system (Escherichia coli). The main kinetic parameters of HIV-1 integrase (K-m = (3.7 +/- 0.2).10(-10) M, k(cat) = (1.2 +/- 0.3).10(-7) sec(-1)) were determined using an oligonucleotide duplex constructed on the basis of the U5-terminal sequence of proviral HIV-1 DNA as the substrate. Inhibition of integrase by aurintricarbonic acid ([I](50) = 6.3 +/- 0.4 muM) and dependence of integrase activity on Mg2+ and Mn2+ concentration were studied.
引用
收藏
页码：988 / 993
页数：6
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