重构新型血管内皮生长因子在毕赤酵母中的表达、纯化与功能鉴定

Expression,Purification and Identification of a Novel Reconstructed VEGF in Pichia Pastroris

血管内皮细胞生长因子为内皮细胞特异的刺激因子,在缺血治疗中具有应用前景.本文通过PCR的方法将VEGF165纤溶酶酶切位点突变并将肝素结合域融合到VEGF165的C末端,且将其进行了酵母表达,并纯化了酵母表达产物,对它的生物活性进行了初步鉴定.结果表明,改构的VEGF可形成蛋白二聚体,且仍保留体外刺激血管生成的能力.VEGF的重构成功与表达产物的获得,为进一步研究其功能奠定了基础.

Vascular endothelial growth factor is a specific stimulator for endothelial cells;it has shown a potential application in tissue ischemia field.This study has muted the heparin sensitive cleavage site and fused the heparin binding domain to the C terminus of VEGF165.Then it carries out yeast expression and purification of the yeast expression product and conducts a preliminary identification of biological activity.The results show that change structure of the reconstructed VEGF protein still able to form protein dimers,and retains the angiogenesis in vitro.The successful expression of reconstructed VEGF expression product has laid the foundation for further research its function.