The Preparation of the Recombinant Fusion Protein Human Retinal Pigment Epithelium-derived Factor and the Analysis of Its Angiogenesis Activity
- VernacularTitle:重组人色素上皮细胞衍生因子融合蛋白的制备及其抑制新生血管活性的研究
- Author:
Ya-Ni WANG
;
Xian-Ning LIU
;
Juan-Li ZHU
;
Na AN
;
Xiu-Ping ZHU
;
- Publication Type:Journal Article
- Keywords:
Human pigment Epithelium-derived factor Neovascularization Prokaryotic expression
- From:
China Biotechnology
2006;0(12):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:The recombinant human retinal pigment epithelium-derived factor(PEDF)protein to be obtained and the angiogenesis of the rPEDF to be identified.Methods: PEDF gene gene was amplified by PCR and cloned into pET32a,rPEDF protein was expressed in E.coli BL21 and confirmed by SDS-PAGE and Western blot.The rPEDF was purified by Ni-NTA on denature condition.The concentration of the rPEDF was determined by Bradford method.The angiogenesis of the rPEDF was determined by chick chorioallantoic membrane(CAM) method.Results: The expression plasmid pET32a-PEDF was constructed successfully.The rPEDF was expressed with stable efficiency in E.coli BL21.The results of the CAM experiment showed that the rPEDF had notable angiogenesis effect in the concentration 0.4、0.04 ng/ml,but had no effect in 4 ng/ml.Conclusion:The PEDF gene was cloned and expressed efficiently,the angiogenesis of the rPEDF to be identified and the activity was worked in certain range.The results can facilitate studying its function and spreading its application.