中文摘要：

在研究HIV-1整合酶（IN）抗药性突变T66I时,发现这一突变同时可以提高整合酶的溶解性。原核表达了IN1-288/T66I和野生型（WT）,取菌体破碎后的上清,SDS-PAGE和his标签蛋白质染色进行分析,结果表明IN1-288/T66I可溶性约是WT的2.4倍。600ml培养基中诱导表达IN1-288/T66I/BL21,亲和层析纯化共收获蛋白质4.72mg。用改进的ELISA方法测定IN1-288/T66I和IN1-288/F185K/C280S链转移催化活性,结果显示两种蛋白质活性基本相当。提供了有别于F185K/C280S突变的另外一种整合酶可溶性表达的途径,IN1-288/T66I重组蛋白还可以应用到整合酶抑制剂筛选中,以获取避开T66I抗药性突变的抑制剂。

英文摘要：

During research of the reported drug-resistance mutation T66I, it was found that this mutation can also improve the solubility of integrase. IN1 -288/T66I and WT were expressed at the same time, 10μl of the supernatant was extracted for SDS-PAGE analysis and His-tagged protein staining. Result shows that the solubility of IN1-288/T66I is 2 - 3 fold of WT. 4.72rag protein was obtained from 600ml of cells expressing IN1-288/T66I. The strand transfer activity of IN1-288/T66I and IN1-288/F185K/C280S were analyzed by advanced ELISA, which shows that two proteins are almost equal in catalytic activity. Another method, being different from F185K / C280S mutation, to improve the soluble expression of integrase has been provided. IN1-288/T66I can also be used to screen integrase inhibitor, which can avoid the drug-resistance brought by T66I mutation.