The Recombinant Expression and Receptor-binding Activity of the B Subunit of Shiga-like Toxin Type Ⅱ / 中国生物工程杂志
China Biotechnology
;
(12)2006.
Article
in Chinese
| WPRIM
| ID: wpr-686066
ABSTRACT
Objective:
To express the B subunit of Shiga-like toxin type Ⅱ,and analyze its expression form and receptor-binding activity.Methods:
The slt2b gene was obtained from EHEC O157∶H7 by PCR,and cloned to the expression vector pET22b(+).The genetically engineered bacteria pET22b(+)-stx2B/BL21 expressed the recombinant StxB after induced with IPTG.The renatured inclusion bodies were purified by ion exchange chromatography.The expression form of rStx2B was investigated by denaturing and native electrophoresis.The receptor-binding activity was confirmed by fluorescence detection and flow cytometer.Result:
The constructed genetically engineered bacteria expressed the rStx2B at a high level.The purified protein was obtained after denaturation,renaturation and ion exchange chromatography.According to the denaturing and native electrophoresis,the rStx2B was expressed in a dimmer form,which consists of two monomers cross linked with disulfide bridge.The rStx2B showed good receptor-binding activity by Hela-binding assay.Conclusion:
The genetically engineered bacteria were constructed successfully.The receptor-binding activity of rStx2B was independent of the pentamers.
Full text:
Available
Index:
WPRIM (Western Pacific)
Language:
Chinese
Journal:
China Biotechnology
Year:
2006
Type:
Article
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