Position: Home > Articles > Cloning and Expression of hly gene from Listeria Monocytogengs
China Dairy Cattle
2008
(11)
2-4
单核细胞增生性李斯特氏菌hly基因的克隆表达
作 者：
刘海瑞;付世新;罗春海
单 位：
黑龙江八一农垦大学动物科技学院
关键词：
单核细胞增多性李斯特氏菌;hly基因;克隆表达
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1 646bp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。
译 名：
Cloning and Expression of hly gene from Listeria Monocytogengs
作 者：
Liu Hairui,Fu Shixin,Luo Chunhai(College of Animal Science and Technology,HLJ August First Land Reclamation University,Daqing 163319)
关键词：
Listeria Monocytogengs;hly gene;Clone expression
摘 要：
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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