摘要
为了研究6×His-tag对来源于痤疮丙酸杆菌的亚油酸异构酶(PAI)可溶性重组表达的影响,我们构建了三种大肠杆菌重组菌株,表达了三种重组PAI:C端带有His-tag亲和标签、N端带有His-tag亲和标签和不加His-tag亲和标签。通过SDS-PAGE和Westernblot分析重组蛋白表达情况,并利用气相色谱检测酶活。实验结果表明:在相似的诱导表达条件及生长状态下,三种大肠杆菌重组菌株均成功表达了具有亚油酸异构酶活性的重组蛋白质,三种重组PAI的表达总量相当,6×His-tag的融合表达促进了PAI蛋白质包涵体的形成,而C端6×His-tag带来的影响最大。
In order to investigate the effect of 6×His-tag on the solubility expression of linoleic acid isomerase（PAI） from Propionibacterium acne in E.coli,recombinant E.coli strains over-expressing PAI,PAIH（PAI fused with C-terminal his-tag）,HPAI（PAI fused with N-terminal his-tag） were constructed,respectively.Recombinant enzymes were analyzed by SDS-PAGE and Western blot.Enzyme activities were verified by gas chromatogram.Under similar growth and inducible expression conditions,three recombinant E.coli strains successfully expressed recombinant proteins which had linoleic acid isomerase activity.The total recombinant protein production levels of three recombinant strains were similar,His-tag had a noticeable negative effect on protein solubility and C-terminal His-tag had a bigger negative effect on solubility of the recombinant protein than N-terminal His-tag.
出处
《食品工业科技》
CAS
CSCD
北大核心
2012年第18期217-220,共4页
Science and Technology of Food Industry
基金
"十二五"国家863计划(2011AA100901)
