重组蛋白F3T7的亚克隆、表达、纯化及生物学活性的初步鉴定

Subcloning,expressing,purifying and primary studing the biological activity of F3T7

目的:克隆编码F3T7的DNA序列,获得融合蛋白F3T7,研究其F3T7的功能。方法:采用亚克隆技术获得原核表达载体PET32a(+)-F3TwM法测定F3T7的抗血管活性。结果:F3T7具有特异结合血管内皮细胞能力,并抑制其增殖和新血管生成。结论:初步表明F3T7仍具特异结合血管内皮细胞,抑制新血管生长的能力。

Objective：To clone the DNA sequence coding the F3TT,generate the fused protein F3TT,and investigate its function. Methods ：We constructed prokaryotic expression vector pET32a （＋）-F3T7 by subcloning technique,then fusion protein F3T7 was expressed and purified by affinity chromatography.and its activity of inhibiting the perliferation of HUVEC was assayed by MTT, then FITC was adopted to mark F3T7 to test its affinity with HUVEC,and its anti-angiogenses was detected by CAM. Results： We primarily certified that the fusion protein F3T7 selectively bound to HUVEC,inhibited its proliferation,and showed anti-angiogenic activity.Conclusion：F3T7 could have the abilities of binding to HUVEC and anti-angiogenesis .