Expression and purification of HIV Integrase in E. col i and  Application in HIV Diagnosis

LIU Lian-xing; HAO Yan-ling; YUAN Guo-liang; XU Bin; CHEN Pei; FU Jing-jing; LIU Yong; SHAO YI-ming

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October 2006

LIU Lian-xing, HAO Yan-ling, YUAN Guo-liang, XU Bin, CHEN Pei, FU Jing-jing, LIU Yong and SHAO YI-ming. Expression and purification of HIV Integrase in E. col i and Application in HIV Diagnosis[J]. Virologica Sinica, 2006, 21(5): 413-416.

Citation: LIU Lian-xing, HAO Yan-ling, YUAN Guo-liang, XU Bin, CHEN Pei, FU Jing-jing, LIU Yong, SHAO YI-ming. Expression and purification of HIV Integrase in E. col i and Application in HIV Diagnosis .VIROLOGICA SINICA, 2006, 21(5) : 413-416.

HIV整合酶在大肠杆菌中的表达纯化及其应用

刘连兴 ^1,2,3 ,
郝彦玲 ² ,
袁国亮 ⁴ ,
许斌 ⁴ ,
傅晶晶 ² ,
刘勇 ² ,
邵一鸣 ^2* ,

1.
1．中国科学院武汉病毒研究所，湖北武汉430071
2.
中国疾病预防控制中心性病艾滋病预防控制中心，北京100050
3.
中国科学院研究生院，北京100049
4.
浙江澳亚生物工程研究院有限公司，浙江杭州310018

摘要

采用pThioHisB系统表达HIV-1整合酶P31蛋白，目的蛋白的表达量达到菌体蛋白的30%左右。通过包涵体洗涤和离子交换层析，蛋白纯度高于95%。用纯化后的P31抗原制备成条带免疫试纸条，检测HIV参考品血清时表现了很好的灵敏度和特异性。本研究为进一步开发HIV-1血清学诊断试剂盒奠定了基础。
- HIV-1
- , 整合酶
- , P31
- , 纯化

Expression and purification of HIV Integrase in E. col i and Application in HIV Diagnosis

LIU Lian-xing ^1,2,3 ,
HAO Yan-ling ² ,
YUAN Guo-liang ⁴ ,
XU Bin ⁴ ,
CHEN Pei ² ,
FU Jing-jing ² ,
LIU Yong ^2* ,
SHAO YI-ming ^1,2*

1.
1. Wuhan Institute of Virology,Chinese Academy of Sciences, Wuhan 430071, China
2.
National Center for AIDS/STD Control and Prevention,Chinese Center for Disease Control and Prevention, Beijing 100050, China
3.
Graduate University of Chinese Academy of Sciences, Beijing 100049, China
4.
Hangzhou Ausia Bi-tech.Co.,Ltd, Hangzhou 310018, China

Abstract

HIV-1 integrase P31 of RL42 strain, a B subtype isolate, was expressed in E.coli using pThioHisB vector(Invitrogen). The recombinant protein represented about 30% of total cellular proteins. and was purified by inclusion body washing and Ion Exchange Chromatography （IEX）. The purity of P31 protein reached more than 95%. Purified P31 antigen displayed fairly good sensitivity and specificity when prepared into immunoblot strips and tested with HIV reference sera. This work established a foundation for the development of HIV detection kits.
- HIV-1
- , Integrase
- , P31
- , Purification

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

Expression and purification of HIV Integrase in E. col i and Application in HIV Diagnosis

1. 1. Wuhan Institute of Virology,Chinese Academy of Sciences, Wuhan 430071, China
2. National Center for AIDS/STD Control and Prevention,Chinese Center for Disease Control and Prevention, Beijing 100050, China
3. Graduate University of Chinese Academy of Sciences, Beijing 100049, China
4. Hangzhou Ausia Bi-tech.Co.,Ltd, Hangzhou 310018, China

Abstract: HIV-1 integrase P31 of RL42 strain, a B subtype isolate, was expressed in E.coli using pThioHisB vector(Invitrogen). The recombinant protein represented about 30% of total cellular proteins. and was purified by inclusion body washing and Ion Exchange Chromatography （IEX）. The purity of P31 protein reached more than 95%. Purified P31 antigen displayed fairly good sensitivity and specificity when prepared into immunoblot strips and tested with HIV reference sera. This work established a foundation for the development of HIV detection kits.