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中国生物工程杂志

China Biotechnology
China Biotechnology  2010, Vol. 30 Issue (08): 17-21    DOI: Q819
    
Research of the Improvement of Soluble Expression of HIV-1 Integrase Induced by T66I Mutation in E.coli
YANG Dong, LIU Bin,HE Hong-qiu,ZHANG Xiao-yi,ZHANG Hai-feng,XU Xian-jin,CHEN Wei-zu,WANG Cun-xin
Life Science and Bioengineering College of Beijing University of Technology , Beijing 100124 ,China
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Abstract  

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. 4.72mg 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. 



Key wordsIntegrase      Soluble expression      T66I mutation     
Received: 17 March 2010      Published: 25 August 2010
Cite this article:

YANG Dong, LIU Bin, HE Gong-Qiu, ZHANG Xiao-Die, ZHANG Hai-Feng, HU Xian-Jin, CHEN Wei-Jie, WANG Cun-Xin. Research of the Improvement of Soluble Expression of HIV-1 Integrase Induced by T66I Mutation in E.coli. China Biotechnology, 2010, 30(08): 17-21.

URL:

https://manu60.magtech.com.cn/biotech/Q819     OR     https://manu60.magtech.com.cn/biotech/Y2010/V30/I08/17

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