›› 2011, Vol. 38 ›› Issue (10): 81-85.

• 生物技术 • Previous Articles     Next Articles

Construction and Expression of Recombinant Baculovirus with P1-2A Gene of Serotype O Foot-and-mouth Disease Virus

LIAO De-fang1, XIN Ai-guo1, GAO Hua-feng1, MIAO Hai-sheng1, GAO Lin1, ZHU Ming-wang2, LI Hua-chun1   

  1. 1. Yunnan Animal Science and Veterinary Institute,Kunming 650224, China;2. Yunnan Baoshan Vaccine Plant,Baoshan 678000, China
  • Received:2011-03-11 Revised:1900-01-01 Online:2011-10-20 Published:2011-10-20

Abstract: Based on the known nucleotide sequence of FMDV gene, P1-2A gene primer was designed and synthesized. P1-2A gene was amplified by RT-PCR. The gene was then cloned into pMD18-T plasmid. The recombinant plasmids were sequenced and cloned into transfer vector pFastBacTMDual. The transfer plasmid pFastBac-P12A was constructed. pFastBac-P12A was further transferred into receptor DH10Bac bacteria containing a shuttle vector Bacmid. The recombinant plasmid Bacmid-P12A was obtained by selection, then was trans-infected into Sf9 insect cells. The recombinant baculovirus which expressing serotype O FMDV P1-2A gene was harvested. The Sf9 cells were trans-infected with recombinant baculovirus expressing serotype O FMDV P1-2A gene and showed typical CPE. The cells were harvested and tested by FMDV Dot blotting and SDS-PAGE analysis. Results indicated that the serotype O FMDV P1-2A gene expressed in Sf9 cells and the P1-2A protein antigen was specific to serotype O FMDV.

Key words: FMDV; P1-2A gene; recombinant baculovirus; expression 

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