Citation: LUO Bao-zheng, BO Qing-ru, CHEN Jin-ding, WANG Wei-yi, CHENG Gang, HE Yun-shao. Expression of FMDV 3ABC Gene in Bac-to-Bac Baculovirus Expression System .VIROLOGICA SINICA, 2005, 20(3) : 303-306.

Expression of FMDV 3ABC Gene in Bac-to-Bac Baculovirus Expression System

  • Available online: 20 June 2005
  • The nonstructural protein gene 3ABC of Foot-and-mouth disease virus (FMDV) was amplified by reverse transcription (RT) PCR and the PCR product was inserted into transfer vector pFastbacHT. Then, the recombinant vector pFastbac HT-3ABC was extracted and transfered into DH-{10}Bac containing a shuttle vector Bacmid. 3ABC gene was integrated into Bacmid by site specific transposition. Subsequently, recombinant shuttle vector Bacmid-3ABC was transfected with Hi Five insect cells. Identification by PCR amplification demonstrated that 3ABC gene was correctly inserted into baculovirus genome at the downstream of polyhedrin promoter. SDS-PAGE and Western blot analysis detected a band of about 50 kDa in the expression product of 3ABC gene in insect cells. The successful expression of 3ABC gene in Bac-to-Bac expression system should provide a new technical platform for the development of novel approaches for distinguishing the infected animals from vaccinated animals.

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    Expression of FMDV 3ABC Gene in Bac-to-Bac Baculovirus Expression System

    • 1. 1. Inst itute of Entomology , Central China Normal University , Wuhan 430079 , China

    Abstract: The nonstructural protein gene 3ABC of Foot-and-mouth disease virus (FMDV) was amplified by reverse transcription (RT) PCR and the PCR product was inserted into transfer vector pFastbacHT. Then, the recombinant vector pFastbac HT-3ABC was extracted and transfered into DH-{10}Bac containing a shuttle vector Bacmid. 3ABC gene was integrated into Bacmid by site specific transposition. Subsequently, recombinant shuttle vector Bacmid-3ABC was transfected with Hi Five insect cells. Identification by PCR amplification demonstrated that 3ABC gene was correctly inserted into baculovirus genome at the downstream of polyhedrin promoter. SDS-PAGE and Western blot analysis detected a band of about 50 kDa in the expression product of 3ABC gene in insect cells. The successful expression of 3ABC gene in Bac-to-Bac expression system should provide a new technical platform for the development of novel approaches for distinguishing the infected animals from vaccinated animals.

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