| 水牛抑制素α亚基基因的克隆与原核表达 |
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| 引用本文： | 谢体三,农微,张新民,黄雅琼,崔奎青,石德顺.水牛抑制素α亚基基因的克隆与原核表达[J].黑龙江畜牧兽医,2009(8). |
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| 作者姓名： | 谢体三 农微 张新民 黄雅琼 崔奎青 石德顺 |
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| 作者单位： | 广西大学,动物繁殖研究所,广西,南宁,530005 |
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| 基金项目： | 广西亚热带生物资源保护利用重点实验室开放课题 |
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| 摘 要： | 采用RT-PCR方法从水牛卵巢总RNA中扩增抑制素α亚基基因,并克隆入pMD18-T载体,进行PCR、双酶切及测序鉴定.序列分析结果表明:水牛抑制素α亚基基因编码序列长为1 083 bp,编码360个氨基酸,与牛、人、猪抑制素α亚基基因CDS成熟蛋白氨基酸的同源性分别为96%、80%、87%,表明抑制素α亚基是一组在进化上高度保守的蛋白质.将水牛抑制素α亚基基因CDS克隆到pET-30a表达载体中,转化宿主菌BL21(DE3)进行原核表达.在1 mmol/L IPTG 中,37 ℃诱导表达4 h后抑制素α亚基基因重组蛋白可成功获得表达.将表达产物进行SDS-PAGE分析,结果表达产物主要以包涵体形式存在,分子质量约为40 ku.
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| 关 键 词： | 抑制素α亚基 水牛 原核表达 |
Cloning and prokaryotic expression of swamp buffalo Inhibin alpha subunit gene |
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| XIE Ti-san,NONG Wei,SHI De-shun,et al.Cloning and prokaryotic expression of swamp buffalo Inhibin alpha subunit gene[J].Heilongjiang Animal Science And veterinary Medicine,2009(8). |
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| Authors: | XIE Ti-san NONG Wei SHI De-shun |
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| Institution: | Animal Reproduction Institute;Guangxi University;Nanning 530005;China |
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| Abstract: | The Inhibin alpha subunit gene was amplified by RT-PCR from total RNA of swamp buffalo ovary and cloned into pMD 18-T vector.The recombinant vector pMD 18-T-INHA was identified by PCR,restriction endonuclease analysis and sequenced.The INHA CDS was amplified and cloned into pET-30a(+) expression vector.The recombinant pET-30a-INHA expression plasmid was confirmed to be constructed successfully by restriction endonuclease analysis and nucleotide sequenced.Then the recombinant plasmid was transformed into BL2... |
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| Keywords: | Inhibin alpha subunit swamp buffalo prokaryotic expression |
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