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水生植物凤眼莲Ca~(2+)-ATPase的原核表达与蛋白纯化; Prokaryotic Expression and Purification of Ca~(2+)-ATPase from Aquatic Plant Eichhornia crassipes
Li YH(李裕红) ; Wu WL(吴文林) ; Zhang N(张鼐)
2014-12-15
关键词凤眼莲 Ca2+-ATPase 原核表达 蛋白纯化 Eichhornia crassipes Ca2+-ATPase prokaryotic expression protein purification
英文摘要从水生植物凤眼莲叶片中提取总rnA,经rT-PCr扩增出CA2+-ATPASE基因片段,经限制性内切酶(SMA I,nOT I)酶切后按正确的读码框顺序插入到PgEX-4T-2表达载体上,重组质粒转化大肠杆菌,经菌落PCr和质粒双酶切鉴定、序列测定确认,证实成功地构建了CA2+-ATPASE基因融合表达载体,.转化菌经IPTg诱导表达,获得了大小约48kd的可溶性目的蛋白,与预期相吻合.利用谷胱甘肽琼脂糖凝胶4b(gluTATHIOnE SEPHArOSE 4b)亲和介质对重组蛋白进行纯化,获得了高纯度的目的蛋白.; The Ca2+-ATPase gene was cloned fromEichhornia crassipes leaves using the PCR technology.After digested by the enzymes(Sma I,Not I),it was inserted into the plasmid pGEX-4T-2to reconstruct the expression vector.The recombinant protein was induced by IPTG and then purified using Glutathione Sepharose 4B.As a result,a single 48 kDa protein was acquired,which implied the protein was the purified Ca2+-ATPase fusion protein.; 国家自然科学基金面上项目(30770391)
语种zh_CN
内容类型期刊论文
源URL[http://dspace.xmu.edu.cn/handle/2288/107038]  
专题化学化工-已发表论文
推荐引用方式
GB/T 7714
李裕红,吴文林,张鼐. 水生植物凤眼莲Ca~(2+)-ATPase的原核表达与蛋白纯化, Prokaryotic Expression and Purification of Ca~(2+)-ATPase from Aquatic Plant Eichhornia crassipes[J],2014.
APA 李裕红,吴文林,&张鼐.(2014).水生植物凤眼莲Ca~(2+)-ATPase的原核表达与蛋白纯化..
MLA 李裕红,et al."水生植物凤眼莲Ca~(2+)-ATPase的原核表达与蛋白纯化".(2014).
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