| 人细胞周期蛋白D1在大肠杆菌中的可溶性表达与纯化 |
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| 引用本文： | 邹德生,张博,张兰杰,辛广,李桂英.人细胞周期蛋白D1在大肠杆菌中的可溶性表达与纯化[J].鞍山师范学院学报,2010,12(2):36-39. |
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| 作者姓名： | 邹德生 张博 张兰杰 辛广 李桂英 |
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| 作者单位： | 鞍山师范学院,化学系,辽宁,鞍山,114007 |
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| 摘 要： | 将人细胞周期蛋白D1基因克隆入原核表达载体pET-20b中获得重组质粒pET-20b-eyeD，经酶切鉴定正确后转化大肠杆菌BL21PlaysS后获得表达菌株．该菌株经IPTG诱导后表达的目的蛋白有部分分泌到培养基上清中，将培养基上清中的蛋白沉淀后用Ni^2＋螯合柱进行纯化，最后可得到纯度达到95％以上的目的蛋白．蛋白电泳显示纯化蛋白的分子量约为33KD，Westemblot分析表明，在电泳胶的相应分子量处出现特异性条带，说明已经成功表达和纯化了重组人细胞周期蛋白D1．
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| 关 键 词： | 人细胞周期蛋白D1 大肠杆菌分泌表达 纯化 大肠杆菌 |
Soluble Expression and Purification of Recombinant Human Cyclin D1 in E.coli BL21 |
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| ZOU De-sheng,ZHANG Bo,ZHANG Lan-jie,XIN Guang,LI Gui-ying.Soluble Expression and Purification of Recombinant Human Cyclin D1 in E.coli BL21[J].Journal of Anshan Teachers College,2010,12(2):36-39. |
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| Authors: | ZOU De-sheng ZHANG Bo ZHANG Lan-jie XIN Guang LI Gui-ying |
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| Institution: | ZOU De-sheng ZHANG Bo ZHANG Lan-jie XIN Guang LI Gui-ying(Department of Chemistry,Anshan Normal University,Anshan Liaoning 114007,China) |
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| Abstract: | The purpose of the study is to obtain soluble expression protein cyclin D1.The recombinant vector pET-20b-cycD is constructed by inserting human cyclin D1 gene into pET-20b plasmid and is identified by digestion with restriction enzymes.An expression strain is selected after transformation of recombined plasmid into E.coli BL21.Part of recombinant protein secrete into culture medium after induced by IPTG.Recombinant protein is purifed by Ni2+ chelate chromatography and the purity of purified protein is up t... |
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| Keywords: | Human cyclin D1 Expression Protein purification E coli BL21 |
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