Soluble expression of a CXCL10-loop3-EGF fusion protein and its anti-tumor activity
- VernacularTitle:融合蛋白CXCL10-loop3- EGF的可溶性表达及其抗肿瘤效应研究
- Author:
Bo SHEN
;
Ying FU
;
Zhefeng MENG
;
Wei XU
;
Xiaofan HE
;
Min ZHU
- Publication Type:Journal Article
- Keywords:
CXCL10-loop3-EGF fusion protein;
Soluble expression;
Antitumor effect
- From:
Chinese Journal of Pathophysiology
2009;25(12):2376-2380
- CountryChina
- Language:Chinese
-
Abstract:
AIM: To evaluate the implication of CXCL10-loop3-EGF fusion protein for the activities of targeting tumor and anti-angiopoiesis. METHODS: RT-PCR was preformed to amplify CXCL10 coding sequence from PBMC activated by IFN-γ. CXCL10-loop3-EGF fusion gene, which was conducted by Over-Lap Extention PCR, was hinged up with plasmid pTG19-T, transfected to E. coli DH5α and processed positive colony selection. After ligated with plasmid pET32a(+), recombinant CXCL10-loop3-EGF fusion gene was then transfected to E. coli Origami B (DE3) and induced to express its coding fusion protein his-CXCL10-loop3-EGF. The recombinant fusion protein CXCL10-loop3 -EGF was purified by His-bind affinity chromatograph, enterokinase cleavage, ultrafiltration and dislysis. The transwell chemotatic test and HUVEC angiopoiesis inhibition test were performed to determine the anti-tumor responses and anti-angiopoiesis activity of CXCL10-loop3-EGF fusion protein. RESULTS: CXCL10-loop3-EGF fusion protein was successfully constructed and confirmed by SDS-PAGE analysis and Western blotting. Significant PBMC chematatic activity and HUVEC anti-angiopoiesis activity were observed. CONCLUSION: CXCL10-loop3-EGF fusion protein, which has perfect anti-tumor activity, is successfully constructed.
