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| 日本脑炎病毒SA14-14-2株E基因的克隆及原核表达 | |
| 引用本文： | 李玲玲,李晨阳,吴阳,周净,王云龙.日本脑炎病毒SA14-14-2株E基因的克隆及原核表达[J].动物医学进展,2007,28(3):23-26. |
| 作者姓名： | 李玲玲 李晨阳 吴阳 周净 王云龙 |
| 作者单位： | 1. 河南省生物工程技术研究中心,河南郑州,450002;河南师范大学生命科学学院,河南新乡,453007 2. 河南省生物工程技术研究中心,河南郑州,450002 3. 河南省生物工程技术研究中心,河南郑州,450002;郑州轻工业学院食品与生物工程学院,河南郑州,450002 |
| 摘 要： | 根据GenBank公布的日本脑炎(Japanese encephalitis virus,JEV)SA14-14-2减毒株E基因的核苷酸序列,设计并合成一对特异性引物,采用RT-PCR方法扩增其E基因全长cDNA,将扩增产物克隆入pUCm-T载体中,测序后亚克隆到原核表达载体pET-32a( ),筛选重组质粒,转化大肠埃希菌BL21(DE3)宿主菌.经IPTG诱导,SDS-PAGE分析表达产物.结果获得了含全长日本脑炎病毒E基因的重组质粒,经测序证实,与GenBank上E基因序列的同源性达到100%.所表达的融合蛋白主要以包涵体形式存在,为制备JEV实验室诊断抗原打下了基础. |
| 关 键 词： | 日本脑炎病毒 减毒活疫苗 E基因 表达 |
| 文章编号： | 1007-5038（2007）03-0023-04 |
| 收稿时间： | 2006-12-18 |
| 修稿时间： | 2006年12月18 |
Cloning and Expression of E Gene of Japanese encephalitis virus(JEV) Strain SA14-14-2 in E.coli |
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| LI Ling-ling,LI Chen-yang,WU Yang,ZHOU Jing,WANG Yun-long.Cloning and Expression of E Gene of Japanese encephalitis virus(JEV) Strain SA14-14-2 in E.coli[J].Progress In Veterinary Medicine,2007,28(3):23-26. | |
| Authors: | LI Ling-ling LI Chen-yang WU Yang ZHOU Jing WANG Yun-long |
| Abstract: | According to the cDNA sequence of JEV E protein derived from the GenBank(Accession No.AF495589),a pair of primers were designed.Genomic RNA was isolated from JEV attenuated strain SA14-14-2,and used as templates for cDNA synthesis of E gene.Subsequently the cDNA encoded JEV E protein was synthesized by using RT-PCR.Then the amplified fragment was cloned into vector pUCm-T and transformed into E.coli TG1.After sequencing,the recombinant expression plasmid pET-32a( )/E was constructed and transformed into E.coli BL21(DE3).After induced by IPTG,SDS-PAGE was performed to detect the E gene product.The E gene had the identies of 100% with the sequence of JEV E protein published in the GenBank(Accession No.AF495589)and the JEV E protein was expressed successfully in inclusion body,which should be useful for the production of diagnostic reagents. |
| Keywords: | Japanese encephalitis virus attenuated vaccine E gene expression |
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