| 人SCG10基因原核质粒构建及其重组蛋白表达 |
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| 引用本文： | 刘芙蓉,李彦妹,张红艳,苏楠,李家滨,李丰.人SCG10基因原核质粒构建及其重组蛋白表达[J].中国医科大学学报,2011,40(6):491-493. |
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| 作者姓名： | 刘芙蓉 李彦妹 张红艳 苏楠 李家滨 李丰 |
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| 作者单位： | 中国医科大学细胞生物学教研室卫生部细胞生物学重点实验室,教育部医学细胞生物学重点实验室,沈阳110001 |
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| 基金项目： | 国家自然科学基金重大研究计划(90813038);国家自然科学基金资助项目(30771128,31000627) |
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| 摘 要： | 目的 构建SCG10的原核表达载体并诱导、纯化和鉴定其表达.方法 pcDNA3.1-SCG10质粒用BamHI和XhoI双酶切后,克隆至pGEX-5X-1栽体,IPTG(异丙基硫代半乳糖苷)在BL21大肠杆茵中诱导glutathione S-transferase(GST)-SCG10 融合蛋白表达,利用GST-beads纯化诱导的融合蛋白,经Western blot印迹鉴定.结果 SCG10全长编码序列克隆至pGEX-5x-1载体中,双酶切鉴定片段大小为500 bp,在BL21大肠杆茵中IPTG诱导融合蛋白的表达分子量为47 kDa,成功纯化出GST及GST-SCGl0蛋白,Wstem blot检测到蛋白表达.结论 构建了SCG10基因原核表达载体,鉴定了GST-SCG10融合蛋白表达.
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| 关 键 词： | SCG10 蛋白纯化 融合蛋白 Western blot |
Construction of Prokaryotic Plasmid of Human SCG10 Gene and Identification of Its Recombinant Protein |
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| LIU Fu-rong
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LI Yan-shu
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ZHANG Hong-yan
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Su Nan
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LI Jia-bin
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LI Feng.Construction of Prokaryotic Plasmid of Human SCG10 Gene and Identification of Its Recombinant Protein[J].Journal of China Medical University,2011,40(6):491-493. |
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| Authors: | LIU Fu-rong LI Yan-shu ZHANG Hong-yan Su Nan LI Jia-bin LI Feng |
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| Institution: | LIU Fu-rong,LI Yan-shu,ZHANG Hong-yan,Su Nan,LI Jia-bin,LI Feng(Department of Cell Biology,China Medical University,Key Laboratory of Cell Biology,Ministry of Public Health,Key Laboratory of Medical Cell Biology,Ministry of Education,Shenyang 110001,China) |
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| Abstract: | Objective To construct prokaryotic expression vector of human SCG10 gene and induce,purify and identify its recombinant protein expression.Methods The coding sequence of SCG10 gene was digested from pcDNA3.1-SCG10 with BamH1 and Xho1 enzymes,and cloned into pGEX-5X-1.The expression of GST-SCG10 fusion protein was induced by IPTG,purified by GST-beads and identified by Western blot.Results The coding sequence of SCG10 gene was cloned into the pGEX-5X-1 vector.The length of the fragment was 500bp.The expressi... |
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| Keywords: | SCG10 protein purification fusion protein Western blot |
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