Cloning of cDNA Encoding Extra-Cellular Domain of Human VEGF Receptor KDR and Effect of Its Recombinant Protein on Endothelial Cell Proliferation and Angiogenesis
- VernacularTitle:VEGF受体KDR克隆及其对内皮细胞增殖和血管新生的影响
- Author:
Caicun ZHOU
;
Lisong TAN
;
Liang TANG
;
Jie ZHANG
;
Bo SU
;
Jian KANG
- Publication Type:Journal Article
- Keywords:
vascular endothelial growth factor;
soluble receptor;
KDR;
procaryotic expression;
pichia yeast;
angiogenesis
- From:
Chinese Journal of Cancer Biotherapy
1995;0(03):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To clone cDNA encoding Ⅴ-Ⅶ extra-cellular domains of VEGF receptor KDR and investigate biological activity of the recombinant protein. Methods: Total RNA from the umbelical venous endothelial cells was obteined. From it, KDR cDNA was cloned into the BamH1 and Kpn 1 sites of pQE31 and made expressed with induction of IPTG. The recombinant protein was purified, refolded and determined for its biological activities. Results: The expression system could express the human recombinant extra-cellular protein of KDR with expression level accounting for 5% of the total bacterial protein. After renaturation, the recombinant protein could bind to 125 I-VEGF and suppress endothelial cell proliferation and angiogenesis of chick chorioallantoic membrance induced by VEGF. Conclusion: Human KDR extra-cellular domain can be expressed in E coli and after being refolded, has an action of binding to VEGF.
