梭鱼白细胞介素-22原核表达及活性蛋白的制备  

Prokaryotic expression and preparation of Liza haematocheila interleukin-22 protein with biological activity

作　　者：王资生[1,2] 齐志涛[1,2] 黄贝[3] 张启焕[1,2] 仇明[1,2] 邵荣[2] 

机构地区：[1]盐城工学院海洋技术系/江苏省沿海池塘养殖生态重点实验室,盐城224051 [2]盐城工学院化学与生物工程学院,盐城224051 [3]中国科学院水生生物研究所/淡水生态与生物技术国家重点实验室,武汉430072

出　　处：《华中农业大学学报》2012年第6期752-755,共4页Journal of Huazhong Agricultural University

基　　金：江苏省自然科学基金项目(BK2011418);江苏省省属高校自然科学基金项目(10KJB240001);盐城工学院人才引进项目(XKR2011007)

摘　　要：根据梭鱼IL-22cDNA序列设计特异性引物,从梭鱼脾脏中扩增IL-22用于表达片段,连接至载体pUC57。分别以BamHⅠ和KpnⅠ对含有目的基因的质粒和pQE-30进行酶切,连接并转化到大肠杆菌M15中,以IPTG进行诱导表达。表达产物通过Ni-NTA亲和层析进行纯化。经SDS-PAGE对表达产物进行鉴定。SDS-PAGE显示该IL-22表达质粒经IPTG诱导后,IL-22重组蛋白主要在菌体培养上清中表达。通过优化蛋白表达及纯化方法,成功获得了梭鱼IL-22活性蛋白。To construct a prokaryotic expression system of Liza haematocheila interleukin（IL）-22,the partial IL-22 gene was cloned from spleen using the specific primers by PCR and inserted into pUC57 vector.Then,the plasmid and pQE-30 vector were digested with BamHⅠ and KpnⅠ,ligated and sub-cloned into M15.The IL-22 protein was expressed by IPTG induction,purified by Ni-NTA and confirmed by SDS-PAGE.The SDS-PAGE result showed that after induced by IPTG,the IL-22 was expressed successfully in M15,and mainly expressed in supernatant.The results indicated that the IL-22 prokaryotic expression vector was constructed successfully and the recombinant protein possessed biological activity.

关 键 词：梭鱼 白细胞介素-22 原核表达 蛋白纯化 

分 类 号：S917.4[农业科学—水产科学]