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


CONCLUSIONS
The shortened HBsAg can be expressed in prokaryocyte.
Ling
Ning
N
Institute for Viral Hepatitis, Chongqing University of Medical Sciences, Chongqing 400010, China.
Ren
Hong
H
Peng
Ming-li
ML
Xu
Hong-mei
HM
Qing
Yu-ling
YL
D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D006514:Hepatitis B Surface Antigens / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology*; D006515:Hepatitis B virus / Q000235:genetics; D006801:Humans; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011387:Prokaryotic Cells / Q000378:metabolism; D011498:Protein Precursors / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology
D006514:Hepatitis B Surface Antigens; D011498:Protein Precursors; D011993:Recombinant Fusion Proteins; C050806:presurface protein 1, hepatitis B surface antigen; C050807:presurface protein 2, hepatitis B surface antigen