The construction and expression of superantigen SEA and antimelanoma ScFv fusion gene / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 750-753, 2003.
Article
in Chinese
| WPRIM
| ID: wpr-249995
ABSTRACT
Two strategies, direct ligation after enzyme digestion and over-lap PCR technology, were adopted to construct a fusion gene which was composed of the antimelanoma single chain antibody gene and the staphylococcal enterotoxin A gene without N-terminal signal sequence. The fusion gene was subcloned into pET28-a vector and transformed into E. coli BL21(DE3). Ni-NTA system was selected to separate and purify the expresstd products. The inhibition ratio of the fusion protein was tested by MTT method. It is shown that the 6His-ScFv-SEA fusion protein can be expressed stably in E. coli BL21 (DE3). The quantity of the fusion protein was shown up to 30% of the total protein of the bacteria and mainly in inclusion body. By activation the effective cells, the fution protein can inhibit the melanoma cell whith expressed corresponding antigen.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Pharmacology
/
Recombinant Fusion Proteins
/
Cell Survival
/
Cells, Cultured
/
Inclusion Bodies
/
Cell Line, Tumor
/
Therapeutic Uses
/
Drug Therapy
/
Electrophoresis, Polyacrylamide Gel
/
Enterotoxins
Limits:
Humans
Language:
Chinese
Journal:
Chinese Journal of Biotechnology
Year:
2003
Type:
Article
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