猪表皮生长因子在大肠杆菌中的串联表达  

Expression of tandem repeats of porcine epidermal growth factor in Escherichia coli

作　　者：贺超 邓璐[1] 汤承[1] 岳华[1] 

机构地区：[1]西南民族大学生命科学与技术学院,四川成都610041

出　　处：《西南民族大学学报（自然科学版）》2015年第2期144-149,共6页Journal of Southwest Minzu University(Natural Science Edition)

基　　金：国家"863计划"课题(2012AA101304);四川省教育厅创新团队(13TD0060)

摘　　要：根据Gen Bank中猪表皮生长因子(p EGF)基因信息设计引物,通过PCR、亚克隆、基因重组等技术,构建得到p EGF拷贝数分别为1、2、3的原核重组表达质粒.用重组质粒转化E.coli BL21,IPTG诱导表达,SDS-PAGE和Western blotting分析结果表明,在37℃诱导条件下,1、2、3拷贝p EGF重组质粒均能表达p EGF蛋白,灰度值分析表明:表达的融合蛋白占菌体蛋白总量分别为9.54%、20.37%、26.79%,表明在原核表达中,对p EGF进行同向串联,增加其拷贝数,可以提高p EGF在表达后融合蛋白中所占比例,相应提高p EGF蛋白产量.本研究为p EGF重组蛋白的高效表达和批量生产提供可靠的依据.Based on the designed primers of porcine epidermal growth factor（pEGF） gene in GenBank, a recombinant prokaryotic expression plasmid with copy numbers as 1,2, and 3 was constructed, by means of PCR, sub-clone, gene recombination and etc. Then the recombinant plasmid was transformed into host cell E. coli BL21 （DE3） ,and induced with IPTG. According to the results of SDS-PAGE and Western blotting, the pEGF protein was expressed by each of the three copies, under induction condition of 37 ℃. As the grey value demonstrates, the expressed fusion protein accocuntes for 9. 54% ,20. 37% and 26. 79 of the total bacterial protein, and the proportion of pEGF in the expressed fusion protein would be enhanced, so as the production of pEGF protein, if connecting pEGF in series and enhancing its copy number. The research offers reliable evidence for the high-efficiency expression of pEGF as well as its mass production.

关 键 词：猪表皮生长因子 亚克隆 诱导表达 

分 类 号：S852.4[农业科学—基础兽医学] S858.28[农业科学—兽医学]