| 猪瘟Erns基因在大肠杆菌中的高效表达 |
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| 引用本文： | 贾洪林,仇华吉,李娜,朱庆虎,罗玉子,童光志. 猪瘟Erns基因在大肠杆菌中的高效表达[J]. 中国预防兽医学报, 2005, 27(3): 161-163 |
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| 作者姓名： | 贾洪林 仇华吉 李娜 朱庆虎 罗玉子 童光志 |
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| 作者单位： | 1. 中国农业科学院,哈尔滨兽医研究所,兽医生物技术国家重点实验室,黑龙江,哈尔滨,150001;延边大学农学院,动物医学系,吉林,龙井,133400
2. 中国农业科学院,哈尔滨兽医研究所,兽医生物技术国家重点实验室,黑龙江,哈尔滨,150001 |
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| 摘 要： | 提取猪瘟病毒石门系强毒株的基因组RNA,用RT_PCR扩增其Erns基因的cDNA,将其克隆到原核表达载体pPROEXTMHTc中,然后转化大肠杆菌DH5α,经IPTG诱导,Erns基因获得高效表达。SDS_PAGE分析显示,表达的重组融合蛋白表现分子量约为31ku。Westernblot分析表明,重组蛋白具有良好的免疫原性。重组蛋白可用于制备诊断抗原,用于标记疫苗接种和野毒感染的鉴别诊断。
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| 关 键 词： | 猪瘟病毒 RT-PCR 原核表达 |
| 文章编号： | 1008-0589(2005)03-0161-03 |
| 修稿时间： | 2004-02-09 |
Expression of classical swine fever virus Erns gene in E. coli |
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| JIA Hong-lin,QIU Hua-ji,LI Na,ZHU Qing-hu,Luo Yu-Zi,TONG Guang-zhi. Expression of classical swine fever virus Erns gene in E. coli[J]. Chinese Journal of Preventive Veterinary Medicine, 2005, 27(3): 161-163 |
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| Authors: | JIA Hong-lin QIU Hua-ji LI Na ZHU Qing-hu Luo Yu-Zi TONG Guang-zhi |
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| Abstract: | The cDNA encoding E~(rns) of classical swine fever virus (CSFV) was amplified by RT_PCR from CSFV genomic RNA,and then cloned into an expression vector pPROEX~(TM)HTc.The resulting recombinant plasmid was named pPROC_E~(rns).A His_tagged recombinant protein was expressed with high efficiency in E.coli transformed by pPROC_E~(rns) after induction with IPTG.The fusion protein has an apparent molecular weight of about 31 ku as indicated by SDS_PAGE analysis,and showed specific immunoreactivity with anti_CSFV sera in Western blotting.The recombinant E~(rns) will be used for establishing differential diagnostic assays accompanying marker or DIVA vaccines. |
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| Keywords: | CSFV RT-PCR prokaryotic expression |
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