Expression and Identification of Single Chain Antibody of Prokaryotic Expression Vector pOPE101-8E5 / 中国生物工程杂志
China Biotechnology
;
(12)2006.
Article
in Chinese
| WPRIM
| ID: wpr-685406
ABSTRACT
In the present study, we inserted the core-streptavidin cDNA into downstream of multi-cloning site of plasmid pOPE101-8E5 by DNA gene recombination technology. And then, the variable fragments of heavy and light chain of the scFv-8E5 were replaced by the scFv-C4 variable fragments to construct the expression vector pOPE101-C4∷core-streptavidin. After transformed the vector pOPE101-C4∷core streptavidin into E.coil, the fusion protein C4∷core streptavidin-His-tag can be expressed by inducing with IPTG, and the expression level and activity of the expressed fusion protein analyzed by SDS-PAGE and Western blot. The results show that a scFv-C4∷core-streptavidin fusion protein of 45kDa was obtained, which can bind proteins of 60kDa & 45kDa from the KG1a cells lysate simultaneously. The binding function can be detected by the binding of core-streptavidin and biotin directly.
Full text:
Available
Index:
WPRIM (Western Pacific)
Type of study:
Diagnostic study
/
Prognostic study
Language:
Chinese
Journal:
China Biotechnology
Year:
2006
Type:
Article
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