Gene expression and activities analysis of a new fusion protein (RGD) 3/tTF / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 409-412, 2007.
Article
in Chinese
| WPRIM
| ID: wpr-328014
ABSTRACT
To develop a new fusion protein (RGD)3/tTF for the therapy of the selective thrombosis of tumor blood vessels. The fused gene (RGD) 3/tTF was reconstructed by PCR, was cloned into vector pET22 b(+), and expressed in E. coli BL21 (DE3). The fusion protein was purified through Nickel-affinity chromatography column. The tTF activity of the fusion protein was detected by clotting assay and F X activation assay. The specific binding of (RGD) 3/tTF to alphavbeta3 was analyzed by indirect ELISA. The recombinant plasmid pET22 b(+)/(RGD)3/tTF was obtained and expressed in E. coli BL21 (DE3). The purified fusion protein could induce blood coagulation, activiate F X. The ability of (RGD) 3/tTF binding specifically to alphavbeta3 was increased by 32%, compared with RGD/tTF. A new fusion protein (RGD) 3/tTF was successfully expressed in E. coli BL21 (DE3). The expressed proteins retained tTF activity and showed a higher binding to alphavbeta3 than that of RGD/tTF.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Oligopeptides
/
Pharmacology
/
Protein Binding
/
Blood Coagulation
/
Recombinant Fusion Proteins
/
Thromboplastin
/
Enzyme-Linked Immunosorbent Assay
/
Factor Xa
/
Gene Expression
/
Polymerase Chain Reaction
Limits:
Animals
/
Humans
Language:
Chinese
Journal:
Chinese Journal of Biotechnology
Year:
2007
Type:
Article
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