| HBsAg和B19 VP2复合抗原的基因克隆及在大肠杆菌中的表达 |
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| 引用本文： | 赵国强,张世杰,刘全离,徐慧丹,陈宗德.HBsAg和B19 VP2复合抗原的基因克隆及在大肠杆菌中的表达[J].郑州大学学报(医学版),2005,40(3):387-390. |
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| 作者姓名： | 赵国强 张世杰 刘全离 徐慧丹 陈宗德 |
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| 作者单位： | 郑州大学基础医学院微生物学与免疫学教研室,郑州,450052;漯河医学专科学校微生物学教研室,漯河,462000;淮阳卫生职业中专微生物学教研室,淮阳,466700 |
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| 基金项目： | 河南省科技攻关基金资助项目 01140512 |
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| 摘 要： | 目的构建乙肝表面抗原(HBsAg)-人微小病毒B19VP2蛋白的原核高效表达系统.方法用PCR扩增HBsAg基因及B19病毒VP2基因,分别与pGEM-T重组,构建出pGEM-T-HBs及pGEM-T-VP2并测序分析;测序证实序列正确后,将pGEM-T-HBs中的HBs基因克隆入pGEX-5X-1表达载体中,得到pGEX-5X-1-HBs;再将pGEM-T-VP2中的VP2基因克隆入pGEX-5X-1-HBs中,得到pGEX-5X-1-HBs-VP2后转化至BL21(DE3)宿主菌中,并以IPTG诱导表达融合蛋白,以Western-blot鉴定.结果pGEX-5X-1-HBs-VP2可在大肠杆菌BL21中高效表达,表达产物经Western-blot鉴定具有HBsAg及B19 VP2的双重抗原性.结论成功构建pGEX-5X-1-HBs-VP2原核表达载体,且其表达的重组蛋白具有良好的抗原性,为HBV和B19病毒重组多价疫苗的研究奠定了基础.
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| 关 键 词： | B19 VP2 HBsAg 原核表达 |
| 修稿时间： | 2005年2月22日 |
Cloning of combining antigen of HBsAg and human parvovirus B19 and its expression in E. Coli. |
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| ZHAO Guoqiang,Zhang Shijie,LIU Quanli,XU Huidan,CHEN Zongde.Cloning of combining antigen of HBsAg and human parvovirus B19 and its expression in E. Coli.[J].Journal of Zhengzhou University: Med Sci,2005,40(3):387-390. |
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| Authors: | ZHAO Guoqiang Zhang Shijie LIU Quanli XU Huidan CHEN Zongde |
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| Institution: | ZHAO Guoqiang 1),ZHANG Shijie 1),LIU Quanli 2),XU Huidan 3),CHEN Zongde 1) 1)Department of Microbiology and Immunology,College of Basic Medical Sciences,Zhengzhou University,Zhengzhou 450052 2)Department of Microbiology,Luohe Medical Technological Academy,Luohe 462000 3)Department of Microbiology,Hygiene and Technical School of Huaiyang,Huaiyang 466700 |
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| Abstract: | Aim: To construct efficient expression vector of HBsAg and B19-VP2 gene. Methods: HBsAg gene and B19 VP2 gene were amplified using PCR, and cloned with pGEM-T vector and sequencing. The HBsAg gene was inserted into the prokaryotic expression vector pGEX-5X-1 and the recombinant pGEX-5X-1-HBs was obtained. Then VP2 gene was inserted into pGEX-5X-1-HBs in order to form pGEX-5X-1-HBs-VP2, which was transformed into E.Coli. BL21(DE3) and induced to express the fusion protein by IPTG. Finally it were confirmed by Western-blot. Results: pGEX-5X-1-HBs-VP2 recombinant vector could express efficiently in BL21. By using Western-blot, it was confirmed that the fusion protein had both the antigen epitopes of HBsAg and B19 VP2 protein partially. Conclusion: The pGEX-5X-1-HBs-VP2 expression vector has been constructed successfully. And its expression product has antigenic characterization of HBsAg and B19 VP2 protein. It provides information for developing B19 and HBsAg recombinant polyvalent vaccine. |
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| Keywords: | B19 VP2 HBsAg prokaryotic expression |
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