中华绒螯蟹卵巢EJO1基因在大肠杆菌BL21(DE3)中的表达  

Expression of Eriocheir sinensis Ovarian EJO1 Gene in E.coli BL21 (DE3)

作　　者：陆钟岩[1] 冯金叶[1] 宋大祥[1] 马长艳[1] 郭豫杰[1] 周开亚[1] 

机构地区：[1]南京师范大学生命科学学院江苏省生物多样性和生物技术重点实验室,南京210093

出　　处：《淡水渔业》2006年第5期3-6,共4页Freshwater Fisheries

基　　金：国家自然科学基金重点项目(No.30130040)

摘　　要：扩增中华绒螯蟹(Eriocheir sinensis)卵巢EJO1基因(GenBank检索号:AY185917)的cDNA全序列得到编码区序列,并将其插入原核表达载体pET28b中,构建成表达质粒pET28b-EJO1。该表达质粒在大肠杆菌(Esche-richia coli)BL21(DE3)中经IPTG诱导表达,得到EJO1-H IS6融合蛋白。SDS-PAGE电泳分析表明,该融合蛋白的相对分子质量约为32 000。经镍离子柱亲和层析进一步纯化得到纯度较高的EJO1-H IS6融合蛋白。EJO1 （GenBank accessing number： AY185917） is an ovarian gene of Chinese mitten crab（Eriocheir sinensis） presumably involved in the ovary development. The complete mature peptide-coding cDNA fragment of EJO1 was obtained by PCR. The product was inserted into the prokaryotic expression vector, pET28b, to produce the expression vector pET28b-EJOI. The recombinant plasmid was transformed into E. coli BL21 （ DE3 ）. EJO1-HIS6 fusion protein was obtained after the addition of IPTG into the growth media. SDS-PAGE analysis revealed that the molecular weight of EJO1- HIS6 fusion protein was about 32 kDa. Purified fusion protein was obtained after purification with NI-NTA His-Bind resin cohlmn.

关 键 词：中华绒螯蟹 EJ01基因 原核表达 EJ01-HIS6融合蛋白 

分 类 号：S966.16[农业科学—水产养殖]