| 斑马鱼促性腺激素释放激素基因的克隆与表达 |
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| 引用本文： | 谢保胜,段瑞君,藤原滋树.斑马鱼促性腺激素释放激素基因的克隆与表达[J].青海医学院学报,2009,30(1):9-13. |
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| 作者姓名： | 谢保胜 段瑞君 藤原滋树 |
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| 作者单位： | 1. 青海大学生物科学系
2. 日本高知大学理学部 |
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| 基金项目： | 青海省高校日元贷款人才培养项目 |
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| 摘 要： | 目的克隆斑马鱼促性腺激素释放激素（GnRH）基因，构建重组质粒pQE30－GnRH并获得表达蛋白。方珐从斑马鱼脑组织提取总RNA，应用RT～PCR方法获得斑马鱼GnRHcD—NA。将该cDNA插入质粒pQE30中，并使其在大肠杆菌RB791中表达。经IPTG诱导后SDS—PAGE电泳检测蛋白的表达情况。结果斑马鱼GnRHcDNA长为207bp，编码的GnRH前体为69个氨基酸残基。与鲤鱼、鲫鱼、拟鲤、黑头软口鲦等淡水鲤科鱼类的氨基酸序列同源性比较，分别为74．4％，69．8％，73．3％和73．3％。电泳结果显示一新的分子量约为14．4kDa的特异蛋白带。结论本研究所克隆的斑马鱼GnRH基因属于GnRH－Ⅱ。构建斑马鱼GnRH原核表达质粒不仅获得大量的GnRH蛋白，还可对该蛋白做进一步分离纯化以及生物活性方面的研究。
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| 关 键 词： | 斑马鱼 促性腺激素释放激素 克隆 基因表达 |
CLONING AND EXPRESSION OF GONADOTROPIN-RELEASING HORMONE GENE FROM ZEABRO FISH(Danio Rerio) |
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| Xie Baosheng,Duan Rui Jun,Shigeki Fujiwara.CLONING AND EXPRESSION OF GONADOTROPIN-RELEASING HORMONE GENE FROM ZEABRO FISH(Danio Rerio)[J].Journal of Qinghai Medical College,2009,30(1):9-13. |
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| Authors: | Xie Baosheng Duan Rui Jun Shigeki Fujiwara |
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| Institution: | 1.The departement of biological science;Qinghai University;2.Faculty of science of Kochi University;Japan |
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| Abstract: | Objective To amplify the gonadotropin-releasing hormone(GnRH) gene in Danio rerio and to construct the recombinant plasmid pQE30-GnRH for obtaining expressed protein.Methods The total RNA was extracted from brain of Danio rerio.The GnRH cDNA was amplified by RT-PCR method.The GnRH cDNA fragment was inserted to plasmid pQE30 and made it expression in E.coli RB791 cells.The expression protein was assayed by SDS-PAGE after IPTG inducing.Results The GnRH cDNA fragment of Danio rerio was 207bp in length and enco... |
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| Keywords: | Zebro fish （Danio rerio） GnRH Clone Gene expression |
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