| 银狐生长激素cDNA克隆和原核表达 |
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| 引用本文： | 刘玉堂,胡晓航,闫丽辉,曹殿军,刘培欣,杨传平.银狐生长激素cDNA克隆和原核表达[J].中国畜牧兽医,2008,35(4):26-30. |
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| 作者姓名： | 刘玉堂 胡晓航 闫丽辉 曹殿军 刘培欣 杨传平 |
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| 作者单位： | 1.东北林业大学，哈尔滨 150040；2.中国农业科学院哈尔滨兽医研究所，哈尔滨 150001；3.黑龙江大学，哈尔滨 150080 |
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| 摘 要： | 为了获得重组的银狐生长激素，本试验用Trizol的方法从银狐脑垂体组织中提取总RNA，以mRNA为模板，用RT-PCR方法扩增出生长激素成熟cDNA片段并克隆到载体质粒pMD18-T中；通过含有BamHⅠ酶切位点的特异引物从pMD18-T-fGH中亚克隆出cDNA片段，并将其重组到pPROEXTMHta质粒中，构建了银狐生长激素原核表达质粒pPROEXTMHta-fGH。将pPROEXTMHta-fGH原核表达质粒转化DH5α大肠杆菌，用终浓度1 mmol/L IPTG进行诱导，通过SDS PAGE进行检测。结果表明在转化的大肠杆菌中检测到分子量为26 kD的fGH蛋白的存在，表达量占菌体蛋白总量的35%，并且表达产物以包涵体的形式存在。
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| 关 键 词： | 银狐 生长激素 克隆 原核表达 |
| 文章编号： | 1671-7236(2008)04-0026-05 |
| 修稿时间： | 2007年11月9日 |
Cloning and Prokaryotie Expression of Growth Hormone cDNA from Vulpes Vulpes |
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| LIU Yu-tang,HU Xiao-hang,YAN Li-hui,CAO Dian-jun,LIU Pei-xin,YANG Chuan-ping.Cloning and Prokaryotie Expression of Growth Hormone cDNA from Vulpes Vulpes[J].China Animal Husbandry & Veterinary Medicine,2008,35(4):26-30. |
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| Authors: | LIU Yu-tang HU Xiao-hang YAN Li-hui CAO Dian-jun LIU Pei-xin YANG Chuan-ping |
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| Institution: | 1.Northeast Forestry University, Harbin 150040, China; 2.Harbin Veterinary Research Institute, CAAS, Harbin 150001, China;3.Heilongjiang University, Harbin 150080, China |
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| Abstract: | To obtain the recombinant vulpes vulpes growth hormone. The mature vulpes vulpes growth hormone mRNA obtained from pituitary was amplified by RT PCR , cloned into vector pMD18-T and subcloned into prokaryotic expression vector pPROEXTMHta at restriction enzyme sites BamHⅠ and HindⅢ. The recombinant plasmids pMD18-TfGH cDNA and pPROEXTMHta-fGH cDNA were identified by restriction enzyme digestion, PCR and sequencing. Then we transfer positive recombinant prokaryotic plasmid into E.Coil, and the protein was expressed under the induction of IPTG at the final concentration of 1 mmol/L. The protein expression was measured by SDS-PAGE, and a high level of fGH protein was obvserved at 26 kD which occupied about 35% of total E.coil protein. The recombined fGH protein existed in the form of inclusion bodies. |
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| Keywords: | vulpes vulpes growth hormone cloning prokaryotic expression |
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