ACTA VETERINARIA ET ZOOTECHNICA SINICA ›› 2010, Vol. 41 ›› Issue (4): 434-441.doi:

• 预防兽医 • Previous Articles     Next Articles

Construction and Screening of the Recombinant Attenuated CPV Expressing P12A3C Gene of FMDV Serotype O

WANG Jianke, ZHANG Yunde, ZHANG Qiang*, WU Guohua, YAN Xinmin,ZHU Haixia, LI Jian, SHAO Changchun, ZHU Caizhu, WU Lei
   

  1. Key Laboratory of Animal Virology of Ministry of Agriculture, State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China
  • Received:2009-02-26 Revised:1900-01-01 Online:2010-04-25 Published:2010-04-25
  • Contact: ZHANG Qiang

PDF (PC)

502

Abstract: The constructed gene EGFPP7.5P12A3C of FMDV O/China99 strain was linked to the linear vector by blunt end ligation using KpnⅠ enzyme site, and the recombinant vector pUC119TKEGFPP7.5P12A3C was obtained. Homologous recombination between recombinant vector with deleted gene TK and capripoxvirus attenuated strain took place in the cell BHK21. The recombinant attenuated strain was screened by choosing EGFP as marker gene. And the recombinant virus was identified by PCR, antigen capture ELISA and Western blot analysis. The recombinant attenuated strain can passage steady in the first to the tenth generation of BHK21 cell, a fragment of about 3 000 bp was amplified，and the gene was confirmed as P12A3C by sequencing. The results of antigen capture ELISA were all positive. Western blot analysis showed that the corresponding protein was expressed in transfer vector pUC119TK EGFPP7.5P12A3C infected GTPV AV41 BHK21 cells and it could be recognized by serotype O of FMDV serum. The result demonstrates that the recombinant attenuated CPV containing P12A3C gene of FMDV serotype O were obtained.