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Development of a high-throughput assay for the HIV-1 integrase disintegration reaction 被引量:8

Development of a high-throughput assay for the HIV-1 integrase disintegration reaction
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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.
出处 《Science China(Life Sciences)》 SCIE CAS 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)
关键词 HIV-1 INTEGRASE IN-CCD DISINTEGRATION HIGH-THROUGHPUT ASSAY HIV-1 integrase IN-CCD disintegration high-throughput assay
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