重组鹅IFN-α蛋白在内含肽-冷激诱导表达系统中的表达  

Expression of recombinant GoIFN-α in the intein-cold shock induced expression system

作　　者：王艺萌[1] 崔子寅[1] 高明春[1] 马波[1] 王君伟[1] 

机构地区：[1]东北农业大学动物医学学院,黑龙江哈尔滨150030

出　　处：《中国兽医科学》2011年第5期508-513,共6页Chinese Veterinary Science

基　　金：黑龙江省留学回国人员基金项目(LC02C08);黑龙江省"十五"攻关计划项目(GB01B503-02)

摘　　要：为了在大肠杆菌系统中高效表达具有抗病毒活性的重组鹅IFN-α(GoIFN-α)蛋白,在GoIFN-α基因上游融合内含肽Ssp Dnabmini-intein(SDI)序列,然后克隆至pET-43.1a(+)载体中,再将获得的pET-43.1a(+)-SDI-GoIFN-α重组载体和pColdⅢ载体分别进行双酶切,构建内含肽-冷激表达载体pColdⅢ-Nu-sA-SDI-GoIFN-α,将重组载体转化至大肠杆菌Rosetta感受态细胞,IPTG低温诱导表达。结果表明,融合基因获得高效表达,表达的融合蛋白主要以可溶形式存在。表达产物经Ni柱纯化后可得到纯度较高的目的蛋白。经CGBQ-GPMV系统检测,证实表达的融合蛋白具有生物学活性,为后续的纯化工作和研究具有天然活性的重组鹅IFN-α奠定了基础。To achieve high-level expression of soluble recombinant GoIFN-α in Escherichia coli, the Ssp Dnabmini-intein（SDI） coding sequence was merged at the upstream of the GoIFN-α cDNA,and then cloned into vector pET43, 1a （＋）. Finally, the recombinant pET-43, 1a（ q-）-SDI-GolFN a plasmid was digested with the restriction enzymes and the small fragment was ligated into the expression vector pColdⅢ predi- gested with the same enzymes to construct the intein-cold shock expression plasmid, named as pCold Ⅲ- SDI-GolFN-α. The recombinant expression plasmids were transformed to E. coli Rosetta and induced by IPTG. The result showed that the fusion protein was expressed in the form of solubility and then the protein was purified through Nit column. The purified protein could be detected by CGBQ-GPMV,indicating that the fusion protein had the antiviral activity. Overall,the successful expression is of significance for further purification of GolFN-α

关 键 词：冷激 内含肽 鹅α干扰素 可溶性表达 

分 类 号：S852.42[农业科学—基础兽医学]