猪脑心肌炎病毒分离株BJC3全长感染性克隆的构建与拯救病毒鉴定

畜牧兽医学报 ›› 2009, Vol. 40 ›› Issue (9): 1350-1357.doi:

猪脑心肌炎病毒分离株BJC3全长感染性克隆的构建与拯救病毒鉴定

张国庆，朱书，盖新娜，郭鑫，陈艳红，查振林，杨汉春*

中国农业大学动物医学院农业部人畜共患病重点开放实验室，北京 100193

收稿日期:2009-02-24 修回日期:1900-01-01 出版日期:2009-09-24 发布日期:2009-09-24
通讯作者: 杨汉春

Construction of Fulllength Infectious Clone for Encephalomyocarditis Virus BJC3 and Identification of Rescued Virus

ZHANG Guoqing，ZHU Shu，GE Xinna，GUO Xin，CHEN Yanhong，

ZHA Zhenlin，YANG Hanchun*

Key Laboratory of Zoonosis of Ministry of Agriculture, College of Veterinary Medicine, China Agricultural University, Beijing 100193, China

Received:2009-02-24 Revised:1900-01-01 Online:2009-09-24 Published:2009-09-24
Contact: YANG Hanchun

摘要/Abstract

摘要：

本研究旨在建立猪脑心肌炎病毒（EMCV）的感染性克隆技术。利用RTPCR分3段扩增出脑心肌炎病毒BJC3株的全基因组cDNA，依次克隆至低拷贝质粒pWSK29，构建出全长质粒pWSKBJC3/w，经体外转录和转染BHK21细胞拯救病毒。结果表明，构建的全长cDNA克隆具有感染性，在BHK21细胞上可拯救出病毒。拯救病毒（命名为rVBJC3W）在BHK21细胞上的生长特性与其亲本病毒BJC3一致，并保持了对小鼠的致病性。本研究成功构建了中国第1株猪脑心肌炎病毒的感染性克隆，为深入研究其分子致病机制提供了必要的工具。

Abstract:

The objective of this study was to construct infectious clone of encephalomyocarditis virus (EMCV). The genomic cDNA of EMCV BJC3 was amplified by three overlapped segments using RTPCR, and cloned into lowcopy plasmid pWSK29 to construct fulllength cDNA clone pWSKBJC3/w. The pWSKBJC3/w was in vitro transcribed and transfected into BHK21 cells to rescue the virus. The results showed that the fulllength cDNA clone was infectious and the virus could be rescued on BHK21 cells. The rescued virus designated rVBJC3W was identified by RTPCR and indirect immunofluorescence assay (IFA). The rescued virus had similar growth characteristics with its parental virus BJC3 and remained the pathogenicity for mice. Our results indicated that the first infectious cDNA clone of EMCV in China was successfully established and provided an essential tool for investigating the molecular basis of pathogenicity of EMCV.

张国庆;朱书;盖新娜;郭鑫;陈艳红;查振林;杨汉春. 猪脑心肌炎病毒分离株BJC3全长感染性克隆的构建与拯救病毒鉴定[J]. 畜牧兽医学报, 2009, 40(9): 1350-1357.

ZHANG Guoqing;ZHU Shu;GE Xinna;GUO Xin;CHEN Yanhong; ZHA Zhenlin;YANG Hanchun. Construction of Fulllength Infectious Clone for Encephalomyocarditis Virus BJC3 and Identification of Rescued Virus[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2009, 40(9): 1350-1357.

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