Scientia Agricultura Sinica ›› 2013, Vol. 46 ›› Issue (1): 187-194.doi: 10.3864/j.issn.0578-1752.2013.01.022

• VETERINARY SCIENCE • Previous Articles     Next Articles

Construction and Rescue of Recombinant Classical Swine Fever Virus with Shimen Structure Protein and Flag Marker

 ZHU  Yuan-Yuan, HAN  Tao, ZOU  Xing-Qi, FAN  Xue-Zheng, XU  Lu, WANG  Qin, ZHAO  Qi-Zu   

  1. Construction and Rescue of Recombinant Classical Swine Fever Virus with Shimen Structure Protein and Flag Marker
  • Received:2012-11-15 Online:2013-01-01 Published:2012-12-09

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Abstract: 【Objective】Vaccination is still the main strategy for CSFV control in China. CSFV HCLV(Hog Cholera Lapinized Virus, HCLV) strain could protect effectively, but it couldn’t be serologically differentiated between infected pigs and vaccinated pigs (DIVA principle), which could been effectively solved by marker vaccine and marker diagnostics.【Method】Based on infectious cDNA clone of CSFV T strain, which is a temperature-sensitive mutation strain, the complete structural protein-coding region of T strain had been replaced by that of ShiMen strain , and the recombinant plasmid SMT was constructed. Additionally, the Flag gene was inserted into C end of E1 protein of the recombinant virus as a positive marker, and the pSMT-Flag was constructed successfully. After electroporation, the recombinant viruses vSMT and vSMT-Flag were passaged by SK6 cell. 【Result】Two recombinant plasmids were identified by specific restriction enzymes，the rescued viruses could be passaged successfully and confirmed by RT-PCR, sequencing and immunoperoxidase monolayer assay(IPMA). 【Conclusion】 This marker virus could react with the Flag specific monoclonal antibody while could not influence the combination between E2 protein and its specific monoclonal antibody, which could be a potential CSF novel marker vaccine.

Key words: classical swine fever , recombinant virus , Flag , marker vaccine

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