VEGF_(121)-KLAK融合蛋白在大肠杆菌BL21(DE3)中的表达及纯化

Expression and Purification of VEGF_(121)-KLAK Fusion Protein in E.coli BL21(DE3)

目的:在大肠杆菌中高效表达并纯化VEGF121与两性分子KLAK的融合蛋白,为进一步研究其抗肿瘤血管形成作用奠定基础。方法:用RT-PCR法扩增目的基因,插入表达载体pET28a后,转化大肠杆菌BL21(DE3),经IPTG诱导表达重组蛋白,对产物进行SDS-PAGE及Western印迹分析。结果:克隆出目的基因,构建了融合蛋白表达载体,诱导表达后经SDS-PAGE检测表明获得了目的条带。结论:在大肠杆菌中高效表达并纯化了融合蛋白VEGF121-KLAK。

Objective： To obtain high-level expression and purification of fusion protein of VEGFI21 and amphoteric molecules KLAK in E.coli BL2,1（DE3）, and laid foundations for studying its effection resistence to neoplastic angiogenesis. Methods： The genes of VEGF121 and KLAK by RT-PCR method was ligated, and was subcloned into pET-28a which was then transformed into competent E.coli BI21（DE3）. Induced by IPTG, VEGFI21-KLAK fusion protein was expressed in recombinant E.coli BL21 （DE3）, and was detected with SDS-PAGE and Western blot. Results： The genes of VEGF121 and KLAK were obtained, and the recombinant expressing vector was constructed. SDS-PAGE and Western blot analysis indicated that the molecular weight of fusion protein was right. Conclusion： The recombinant expressing vector constructed could produce VEGF121-KLAK fusion protein, and the high purified fusion protein was obtained.