[缩短型乙型肝炎表面抗原的原核表达及抗原性分析]

[Prokaryocytic expression of the shortened hepatitis B surface antigen and antigenic analysis].

作者信息

Ling Ning, Ren Hong, Peng Ming-li, Xu Hong-mei, Qing Yu-ling

机构信息

Institute for Viral Hepatitis, Chongqing University of Medical Sciences, Chongqing 400010, China.

出版信息

Zhonghua Gan Zang Bing Za Zhi. 2003 Apr;11(4):209-11.

PMID:12716516

Abstract

OBJECTIVE

To study the expression of the shortened hepatitis B surface antigen in prokaryocyte and detect the antigenic characters.

METHODS

Firstly, the gene fragments coding the 152 and 124 amino acids of the carboxyl terminus of hepatitis B surface antigen (HBsAg) were amplified by polymerase chain reaction (PCR). Secondly, they were cloned to plasmid pBKS+, and the accuracy of those constructions were confirmed by restriction enzyme digestion and DNA sequencing. Then, they were cloned to prokaryocytic expression vector-plasmid pET32a(+). The recombinant plasmids were transfected into E.coli BL21 and induced to express with IPTG.

RESULTS

The recombinant plasmids were successfully constructed. In E.coli BL21, the protein was expressed in a fusion fashion and could be recognized by monoclonal antibody against HBsAg with ELISA and Western blot.

CONCLUSION

The shortened HBsAg can be expressed in prokaryocyte.

摘要

目的

研究乙型肝炎表面抗原截短体在原核细胞中的表达并检测其抗原特性。

方法

首先，通过聚合酶链反应（PCR）扩增编码乙型肝炎表面抗原（HBsAg）羧基末端152和124个氨基酸的基因片段。其次，将它们克隆到质粒pBKS +中，并通过限制性内切酶消化和DNA测序确认这些构建体的准确性。然后，将它们克隆到原核表达载体-质粒pET32a(+)中。将重组质粒转染到大肠杆菌BL21中并用异丙基-β-D-硫代半乳糖苷（IPTG）诱导表达。