His-猪生长激素融合基因在大肠杆菌中的表达与纯化  

Expression and Purification of His-Porcine Growth Hormone Fusiongene in E.coli

作　　者：朱江[1] 曹广力[1] 姜秀英[2] 王雪峰[1] 朱艳[1] 邓忠斌[1] 李新平[1] 沈颂东[1] 马泓冰[1] 贡成良[3] 

机构地区：[1]苏州大学生命科学学院,苏州215006 [2]江苏徐州医药学校,徐州221116 [3]苏州大学农业科学与技术学院,苏州215006

出　　处：《科技通报》2005年第1期63-68,共6页Bulletin of Science and Technology

基　　金：江苏省教育厅自然科学基金(编号Q1114020)

摘　　要：将通过PCR技术改建并去除了信号肽的猪生长激素cDNA克隆入大肠杆菌表达载体pET-28a(+),构建出能利用Ni-NTAAgarose柱一步纯化表达产物的重组质粒pPGH020。探讨了大肠杆菌中重组猪生长激素的最佳表达条件和纯化方法,制备并纯化了兔抗pGH抗体。SDS鄄PAGE和Westernblot的分析结果表明,26.5kD处有包括His·Taq、thrombin位点序列和pGH序列的融合蛋白特异条带。凝胶扫描估测猪生长激素的表达量占大肠杆菌胞浆蛋白的17.3%左右。The cDNA of pGH without signal peptides was reconstructed by PCR, and then recombined with the vector pET-28a(+) in E.coli. The recombined plasmid, pPGH020 could be directly purified by Ni-NTA Agarose. The best conditions of expression and purification were studied. SDS-PAGE and Western blot analysis showed that the pGH gene product, 26.5kD fusion protein consisted of His·Taq, thrombin cut site and pGH sequence. Gel scanning showed that the expressional quantity of pGH in E.coli were about 17.3 % percents of the total proteins.

关 键 词：猪生长激素基因 大肠杆菌表达 纯化 抗体IGG 

分 类 号：Q784[生物学—分子生物学] Q786