利用大肠杆菌表达系统制备抗VEGF螺旋-环-螺旋多肽及其纯化的研究  

Study on Preparation and Purification of Anti-VEGF Helix-loop-helix Peptide Using Escherichia coli Expression System

作　　者：庹有朋 王刚[1] 叶正茂 藤井郁雄 万玉军[1] 罗丽娟 李楠臻 岳晓敏[1] Tuo Youpeng;Wang Gang;Ye Zhengmao;Ikuo Fujii;Wan Yujun;Luo Lijuan;Li Nanzheng;Yue Xiaomin(Institute of Biological Fermentation,Sichuan Food Fermentation Industry Research and Design Institute,Chengdu 611130,China;Department of Biological Science,Graduate School of Science,Osaka Prefecture University,Osaka,599-8531,Japan)

机构地区：[1]四川省食品发酵工业研究设计院生物发酵研究所,成都611130 [2]大阪府立大学研究生院生物科学系,大阪府,599-8531,日本

出　　处：《基因组学与应用生物学》2021年第1期224-229,共6页Genomics and Applied Biology

基　　金：四川省科技计划项目(2018HH0060);四川省食品防腐保鲜剂微生物发酵技术工程实验室资助。

摘　　要：VEGF (血管内皮生长因子)是一种诱导肿瘤血管形成的作用最强、特异性最高的血管生长因子。本实验利用抗VEGF螺旋-环-螺旋多肽的基因M49,通过PCR技术扩增获得目的基因,构建原核表达pET-32a(+),转入大肠杆菌(Escherichia coli) BL21进行表达,并对目的蛋白进行诱导表达及纯化。结果表明PCR扩增得到186 bp基因片段,诱导表达后得到一个约为22 kD融合蛋白,并进一步使用SUMO蛋白酶切除标签,得到约3.5 kD目的蛋白,通过小鼠(Mus musculus)给药实验验证了目的蛋白的功能性。这将为下一步VEGF螺旋-环-螺旋多肽靶向药的研制提供了必要的条件。Vascular endothelial growth factor(VEGF) is the most powerful and highly specific factor for vascular growth that can induce the tumor angiogenesis. In this experiment, anti-VEGF helix-loop-helix polypeptide gene M49 was amplified by PCR to get the target gene. Then the prokaryotic expression plasmid with vector pET-32 a(+) was constructed and transformed into Escherichia coli The expression of target gene was induced and the products were purified. The results showed that 186 bp gene fragments were obtained by PCR and about 22 kD of fusion protein was obtained after the inducible expression. Cleavaged the tag by SUMO protease, approximately 3.5 kD of the target protein was obtained. The administration experiment in mice confirmed the function of the target protein, so as to provide necessary conditions for the further study and development of the target drug that is anti-VEGF helix-loop-helix polypeptide.

关 键 词：抗VEGF多肽 大肠杆菌表达系统 多肽纯化 

分 类 号：Q78[生物学—分子生物学]