| HIV—1 P24截短体与gp41截短体的融合蛋白在大肠杆菌中的表达、纯化和鉴定 |
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| 引用本文： | 陈伟,闭兰,朱华松,李茜,余模松.HIV—1 P24截短体与gp41截短体的融合蛋白在大肠杆菌中的表达、纯化和鉴定[J].微生物学免疫学进展,2001,29(4):1-5. |
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| 作者姓名： | 陈伟 闭兰 朱华松 李茜 余模松 |
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| 作者单位： | 卫生部武汉生物制品研究所,武汉,430060 |
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| 摘 要： | 用基因重组技术将截短的HIV－1 p24基因和gp41基因连接成嵌合基因，插入质粒pGEX-4T3,构建成重组表达质粒pGEX-F。将pGEX-F转化大肠杆菌BL21。经IPTG诱导表达，pGEX-F在大肠杆菌BL21中获得了高效表达。融合蛋白P24－gp41经Glutathione-Sepharose4B亲和层析纯化后，用间接ELISA和免疫印迹检测HIV抗体阳性血清和正常人血清，P24－gp41只与HIV抗体阳性血清反应，证明获得的融合蛋白P24－gp41有很强的抗原特异性和免疫反应性，具有较高的应用价值。
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| 关 键 词： | HIV－1 P24 gp41 融合蛋白 大肠杆菌 表达 纯化 截短体 免疫诊断试剂 |
| 文章编号： | 1005-5673(2001)-04-0001-05 |
| 修稿时间： | 2001年4月9日 |
Expression,purification and characterization of HIV-1 recombinant P24-gp41 fusion protein in E.coli |
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| CHEN Wei,BI Lan,ZHU Hua Song,et al..Expression,purification and characterization of HIV-1 recombinant P24-gp41 fusion protein in E.coli[J].Progress In Microbiology and Immunology,2001,29(4):1-5. |
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| Authors: | CHEN Wei BI Lan ZHU Hua Song |
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| Abstract: | To prepare a multivalent antigen of HIV 1 for diagnostic reagent,a recombinant expression plasmid pGEX F was constructed by inserting a chimeric gene encoding a fusion protein p24 gp41 of HIV 1 into plasmid pGEX 4T3.The pGEX F was trasfered into E.coli(BL 21 )and expressed by IPTG induction.The recombinant fusion P24 gp41 was purified by Glutathione Sepharose 4B affinity chromatography and characterized with indirect ELISA and Western blot.The recombinant fusion protein P24 gp41 was highly expressed in E.coli(BL 21 )and the purified P24 gp41 only reacted with HIV antibody positive serum in above tests.The recombinant P24 gp41 has excellent antigenigcity and immunoreactivity in relation to prosperous application. |
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| Keywords: | HIV 1 P24 gp41 Fusion protein E coli Expression Purification |
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