摘要
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.
出处
《生物技术通讯》
CAS
2006年第2期170-173,共4页
Letters in Biotechnology