Prokaryocytic expression of the shortened hepatitis B surface antigen and antigenic analysis.
- Author:
Ning LING
1
;
Hong REN
;
Ming-li PENG
;
Hong-mei XU
;
Yu-ling QING
Author Information
- Publication Type:Journal Article
- MeSH: Cloning, Molecular; Escherichia coli; genetics; metabolism; Hepatitis B Surface Antigens; biosynthesis; genetics; immunology; Hepatitis B virus; genetics; Humans; Plasmids; Polymerase Chain Reaction; Prokaryotic Cells; metabolism; Protein Precursors; biosynthesis; genetics; immunology; Recombinant Fusion Proteins; biosynthesis; genetics; immunology
- From: Chinese Journal of Hepatology 2003;11(4):209-211
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo study the expression of the shortened hepatitis B surface antigen in prokaryocyte and detect the antigenic characters.
METHODSFirstly, 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.
RESULTSThe 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.
CONCLUSIONThe shortened HBsAg can be expressed in prokaryocyte.
