ACTA VETERINARIA ET ZOOTECHNICA SINICA ›› 2009, Vol. 40 ›› Issue (9): 1350-1357.

• 预防兽医 • Previous Articles     Next Articles

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*

   

  1. 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

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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.

Key words: encephalomyocarditis virus (EMCV), infectious cDNA clone, virus rescue, growth characteristics, pathogenicity