Preparation and identification of polyclonal antibody against methyl-accepting chemotaxis signal transduction protein of Helicobacter hepaticus.
- Author:
Shengping HE
1
;
Yahua CHEN
;
Yingying ZHAO
;
Jide WANG
;
Yang BAI
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Antibodies, Bacterial; biosynthesis; genetics; Antibody Specificity; Bacterial Proteins; immunology; Helicobacter hepaticus; immunology; metabolism; Male; Mice; Mice, Inbred BALB C; Rabbits; Recombinant Fusion Proteins; biosynthesis; genetics; Signal Transduction
- From: Journal of Southern Medical University 2013;33(9):1295-1298
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo prepare the polyclonal antibody against methyl-accepting chemotaxis signal transduction protein (MCP) of Helicobacter hepaticus (H.hepaticus).
METHODSThe recombinant plasmid pET22b+/MCP was transformed into E.coli BL2l(DE3) to express the fusion protein His-rhMCP under the induction of IPTG. The fusion protein was purified and the antibody was obtained by immunizing rabbits. The titer of the polyclonal antibody was tested by indirect ELISA, and the specificity of the antibody was identified based on Western blotting using the prepared cell surface proteins (CSPs) of the bacteria.
RESULTSThe fusion protein was successfully expressed, and the titer of the antibody reached 1:32 000. Western blotting indicated that the antibody could specifically bind to CSPs and His-rhMCP.
CONCLUSIONThe antibody with a high titer and specificity was prepared to facilitate further study of the pathogenicity and epidemiology of H.hepaticus in human.
