| 河南华溪蟹金属硫蛋白原核表达载体的构建 |
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| 引用本文： | 张志明,马文丽,李涌泉,王兰.河南华溪蟹金属硫蛋白原核表达载体的构建[J].安徽农业科学,2011,39(26):16132-16133. |
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| 作者姓名： | 张志明 马文丽 李涌泉 王兰 |
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| 作者单位： | 山西大学生命科学学院,山西太原,030006 |
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| 基金项目： | 国家自然科学基金项目(30870267与30970361); 山西省自然科学基金项目(2008011069); 山西省回国留学人员科研项目 |
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| 摘 要： | 目的]构建河南华溪蟹（Sinopotamon henanense）金属硫蛋白（MT）的原核表达载体。方法]以大肠杆菌DH-5α中的pGEM-MT质粒为模板,PCR扩增目的片段并连接至pGEM-T载体。然后,亚克隆至pQE31表达载体,转化大肠杆菌DH-5α感受态细胞。结果]PCR扩增目的片段产物大小约为220 bp,与预期大小一致。重组质粒pQE31-MT双酶切鉴定结果也与预期相符。结论]成功构建了MT原核表达载体,为今后表达并纯化MT提供了材料。
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| 关 键 词： | 金属硫蛋白 基因重组 表达载体pQE31 原核表达 |
Construction of Prokaryotic Expression Vector for Metallothionein of Sinopotamon henanense |
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| ZHANG Zhi-ming et al.Construction of Prokaryotic Expression Vector for Metallothionein of Sinopotamon henanense[J].Journal of Anhui Agricultural Sciences,2011,39(26):16132-16133. |
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| Authors: | ZHANG Zhi-ming |
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| Institution: | ZHANG Zhi-ming et al(School of Life Science,Shanxi University,Taiyuan,Shanxi 030006) |
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| Abstract: | Objective] To construct the prokaryotic expression vector for metallothionein(MT) of Sinopotamon henanense.Method] The plasmid pGEM-MT from E.coli DH-5α was used as the template.The target fragment of MT gene was amplified using PCR,and then bound to the vector pGEM-MT for amplification.The amplified fragment of the MT gene was subcloned into the prokaryotic expression vector pQE31 and then transferred to E.coli DH-5α.Result] DNA fragment in size of 220 bp were amplified.The size of the PCR products met our expectation and the results of recombinant plasmid pQE31-MT double-enzyme cleavage was in line with the expectation.Conclusion] A prokaryotic expression vector for MT has been successfully constructed,provided materials for the expression and the purification of the protein. |
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| Keywords: | Metallothionein Gene recombination Expression vector pQE31 Prokaryotic expression |
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