Development of a high-throughput assay for the HIV-1 integrase disintegration reaction  

作　　者：HE HongQiu,LIU Bin,ZHANG XiaoYi,CHEN WeiZu & WANG CunXin College of Life Science and Bioengineering,Beijing University of Technology,Beijing 100124,China 

出　　处：《Science China(Life Sciences)》2010年第2期241-247,共7页中国科学（生命科学英文版）

基　　金：supported by the National Natural Science Foundation of China (Grant No. 30670497);the Beijing Natural Science Foundation (Grant No. 5072002)

摘　　要：Both HIV-1 integrase (IN) and the central catalytic domain of IN (IN-CCD) catalyze the disintegration reaction in vitro.In this study,IN and IN-CCD proteins were expressed and purified,and a high-throughput format enzyme-linked immunosorbent assay (ELISA) was developed for the disintegration reaction.IN exhibited a marked preference for Mn2+ over Mg2+ as the divalent cation cofactor in disintegration.Baicalein,a known IN inhibitor,was found to be an IN-CCD inhibitor.The assay is sensitive and specific for the study of disintegration reaction as well as for the in vitro identification of antiviral drugs targeting IN,especially targeting IN-CCD.Both HIV-1 integrase (IN) and the central catalytic domain of IN (IN-CCD) catalyze the disintegration reaction in vitro.In this study,IN and IN-CCD proteins were expressed and purified,and a high-throughput format enzyme-linked immunosorbent assay (ELISA) was developed for the disintegration reaction.IN exhibited a marked preference for Mn2+ over Mg2+ as the divalent cation cofactor in disintegration.Baicalein,a known IN inhibitor,was found to be an IN-CCD inhibitor.The assay is sensitive and specific for the study of disintegration reaction as well as for the in vitro identification of antiviral drugs targeting IN,especially targeting IN-CCD.

关 键 词：HIV-1 INTEGRASE IN-CCD DISINTEGRATION HIGH-THROUGHPUT ASSAY 

分 类 号：R512.91[医药卫生—内科学]