| 大鼠生精相关基因TSARG1原核表达载体的构建和表达 |
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| 引用本文： | 张懿,刘刚.大鼠生精相关基因TSARG1原核表达载体的构建和表达[J].实用预防医学,2011,18(8):1387-1389. |
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| 作者姓名： | 张懿 刘刚 |
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| 作者单位： | 1. 湖南省长沙市第八医院,湖南,长沙,410100
2. 中南大学生殖与干细胞工程研究所 |
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| 基金项目： | 国家自然科学基金资助项目 |
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| 摘 要： | 目的构建大鼠生精相关基因pGEX-KG/TSARG1重组载体并进行原核表达。方法应用RT-PCR技术从大鼠睾丸组织mRNA中扩增TSARG1的开放阅读框（ORF）,T-A克隆后将TSARG1插入到原核表达载体pGEX-KG中,经测序鉴定后将重组质粒转入宿主菌E.coliBL21,IPTG诱导表达GST/TSARG1融合蛋白。结果成功构建pGEX-KG/TSARG1重组质粒;转化重组质粒的E.coli BL21经IPTG 37℃诱导高效表达GST/TSARG1融合蛋白。结论 pGEX-KG/TSARG1原核表达载体的成功构建为进一步研究TSARG1的生物学功能奠定了基础。
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| 关 键 词： | TSARG1 基因克隆 蛋白表达 精子发生 |
Construction and Expression of pGEX-KG/TSARG1 Recombinant Vector in Rats |
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| ZHANG Yi,LIU Gang.Construction and Expression of pGEX-KG/TSARG1 Recombinant Vector in Rats[J].Practical Preventive Medicine,2011,18(8):1387-1389. |
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| Authors: | ZHANG Yi LIU Gang |
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| Institution: | ZHANG Yi,LIU Gang(The Eighth Hospital of Changsha,Changsha 410100,Hunan,China) |
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| Abstract: | Objective To construct pGEX-KG/TSARG1 recombinant vector in rats.Methods The open reading frame(ORF) of TSARG1 was amplified from rats' testis RNA by RT-PCR.The products were cloned into pUCm-T vectors and then sequenced.Then the recombinant plasmid was digested and subcloned into pGEX-KG vector.The recombinant plasmids were identified by DNA sequence analysis and transformed into component E.coli BL21 cells,the GST/ TSARG1 fusion protein was expressed with IPTG induction.Results The pGEX-KG/TSARG1 recombin... |
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| Keywords: | TSARG1 |
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