【摘要】 目的:建立一种能够较好地分析HIV-1整合酶的生物学活性以及大规模地筛选整合酶抑制剂的方法。方法:HIV-1整合酶经表达复性及纯化后,采用一种基于Biotin-Avidin EILSA(BA-ELISA)的方法,使用化学发光底物测定分析整合酶的生物学活性,以及型核糖体失活蛋白luffin-a对整合酶的抑制作用。结果:1采用BA-ELISA法得到整合酶的比活性为54.92U/mg蛋白;2luffin-a对整合酶的半数抑制浓度为(0.63±0.026)μmol/L。结论:本实验所采用的基于BA-ELISA的方法能够较好地分析HIV-1整合酶的生物活性,适合于大规模体外HIV-1整合酶抑制剂的筛选。更多还原

【Abstract】 Objective: To develop an ELISA-based method for analyzing biologic activities of HIV-1 integrase and for high throughput screening of integrase inhibitors. Methods: After expression,renaturation and purification of integrase,the bioactivity of integrase and the inhibition of luffin-a were evaluated with an in vitro assay based on biotin-avidin EILSA and chemiluminescent substrates. Results: (1)The specific activity of the purified integrase was 54.92 units/mg of protein.(2)IC50 (concentration causing 50% inhibition of integrase) of luffin-a was (0.63±0.026)μmol/L. Conclusion: The non-radioactive assay can be used for analysis of bioactivities and high throughput screening of inhibitors of HIV-1 integrase.更多还原