| 单核细胞增生性李斯特氏菌hly基因的克隆表达 |
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| 引用本文： | 刘海瑞,付世新,罗春海.单核细胞增生性李斯特氏菌hly基因的克隆表达[J].中国奶牛,2008(11):2-4. |
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| 作者姓名： | 刘海瑞 付世新 罗春海 |
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| 作者单位： | 黑龙江八一农垦大学动物科技学院,大庆,163319 |
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| 基金项目： | 黑龙江省大庆市科技局资助项目
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| 摘 要： | 以单核细胞增多性李斯特氏菌（Listeria Monocytogengs，LMO）LMO-0586基因组DNA为模板，运用PCR扩增得到片段大小为1646bp的致病基因李氏溶血素（hly）基因，并克隆到pMD18-T载体上，构建克隆载体pMD18-T-hly。经测序正确后，将hly基因克隆至表达载体pGEX上构建表达质粒pGEX-6P-1-hly。在IPTG诱导下，携带pGEX-6P-1-hlyA的E．coli BL21（DE3）高效表达分子量约为82KDa的可溶性蛋白及包涵体形式的蛋白。为进一步研究溶血素蛋白的结构、功能，LMO的分子流行病学调查、诊断试剂盒的开发提供依据。
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| 关 键 词： | 单核细胞增多性李斯特氏菌 hly基因 克隆表达 |
Cloning and Expression of hly gene from Listeria Monocytogengs |
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| Liu Hairui,Fu Shixin,Luo Chunhai.Cloning and Expression of hly gene from Listeria Monocytogengs[J].China Dairy Cattle,2008(11):2-4. |
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| Authors: | Liu Hairui Fu Shixin Luo Chunhai |
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| Institution: | Liu Hairui, Fu Shixin, Luo Chunhai (College of Animal Science and Technology, HLJ August First Land Reclamation University, Daqing 163319) |
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| Abstract: | The hly gene was amplified by PCR with the genomic DNA of Listeria Monocytogengs LMO-0586 as the template.The hly gene was inserted in pMD18-T to construct the recombinant cloning vector pMD-hly.After the DNA sequence was determined,the hlyA was subcloned into expression vector pGEX to construct the recombinant expression vector pGEX-6P-1-hly.Upon IPTG induction,soluble hly was over-produced by E.coli BL21(DE3) harboring the expression construct.Recombinant showed a single band about 82 KDa on SDS-PAGE gel. |
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| Keywords: | Listeria Monocytogengs hly gene Clone expression |
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