中国药理学通报Issue(10)：1469-1473,5.DOI:10.3969/j.issn.1001-1978.2014.10.029
HIV-1整合酶3′端加工抑制剂筛选方法的建立与优化
Establishment and optimization of a method for screening HIV-1 integrase 3′-processing inhibitor
摘要
Abstract
Aim To establish and optimize a method for screening HIV-1 integrase 3′-processing inhibitor. Methods Fluorescence resonance energy transfer ( FRET) was used to create an assay for screening in-tegrase 3′-processing inhibitors; wavelength was de-fined by DNaseⅠ; factors affecting IN activity were optimized, including buffer composition, substrate con-centration, enzyme concentration, metal ion concentra-tion. Results Integrase 3′-processing optimizing reac-tion conditions were buffer 1 , 500 nmol · L-1 sub-strate, 1 μmol·L-1 integrase, 20mmol·L-1 magne-sium ion. Positive drug raltegravir and myricetin could effectively inhibit integrase 3′-processing activity using this assay. Two integrase 3′-processing inhibitors were screened by this method. Conclusion The method for screening HIV-1 integrase 3′-processing inhibitor is successfully established and optimized.关键词
HIV-1/整合酶/3′端加工/荧光共振能量转移/影响因素/优化/抑制剂Key words
HIV-1/integrase/3′-processing/FRET/influence factors/optimization/inhibitor分类
医药卫生引用本文复制引用
陆翠林,张旋,詹金彪,杨柳萌,郑永唐..HIV-1整合酶3′端加工抑制剂筛选方法的建立与优化[J].中国药理学通报,2014,(10):1469-1473,5.基金项目
国家自然科学基金资助项目( No 81102483) （ No 81102483）
国家科技部“重大新药创制”重大专项课题( No 2009ZX09501-029,2012ZX10001-006,2013ZX09103001-010) （ No 2009ZX09501-029,2012ZX10001-006,2013ZX09103001-010）
