Fusion expression of human renal cell carcinoma-associated antigen G250/MN/CA IX in prokaryotic expression system.
- Author:
Yao-dong JIANG
1
;
Shao-bin ZHENG
;
Wang-long TAN
;
Shan-chao ZHAO
;
Fei REN
;
Bao ZHANG
Author Information
- Publication Type:Journal Article
- MeSH: Antibodies, Monoclonal; immunology; Antibody Specificity; immunology; Antigens, Neoplasm; genetics; immunology; metabolism; Biomarkers, Tumor; genetics; immunology; metabolism; Blotting, Western; Carbonic Anhydrase IX; Carbonic Anhydrases; genetics; immunology; metabolism; Cloning, Molecular; Escherichia coli; genetics; Gene Expression; Humans; Recombinant Fusion Proteins; genetics; immunology; metabolism
- From: Journal of Southern Medical University 2007;27(3):307-309
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo achieve high expression of human renal cell carcinoma-associated antigen G250 in Escherichia coli.
METHODSThe gene fragments encoding the protein obtained by PCR was cloned into prokaryotic expression vector pET32a(+) and expressed in E. coli Rosseta. The immunogenicity of the recombinant protein was evaluated by Western blotting.
RESULTSThe plasmid pET32a(+)/G250 was constructed and expressed in E. coli Rosseta successfully. Western blot analysis showed that the recombinant protein could be specifically recognized by monoclonal antibody M75.
CONCLUSIONEfficient G250 expression is achieved in prokaryotic expression system, which may facilitate further functional study of the protein and its monoclonal antibody preparation.
