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Citation: XU Lin, GUO Yan, GUAN Jie, YU Hong, ZHOU Yu-sen, LI Jin-song*. Construction of Recombinant Adenovirus Expressing Protective Antigen of Bacillus Anthracis and Analysis of Immuno-responses [J].VIROLOGICA SINICA, 2006, 21(3) : 231-234.

Construction of Recombinant Adenovirus Expressing Protective Antigen of Bacillus Anthracis and Analysis of Immuno-responses

  • In this study, we constructed recombinant adenovirus containing protective antigen gene of Bacillus anthracis. The PA gene was obtained from pcDNA3.1-PA vector by PCR , and cloned into plasmid pAdTrack-CMV. The positive clone was linearized with PmeI restriction enzyme and transformed into E. coli BJ5183 cells with the adenoviral plasmid pAdEasy-1. Then vAd-PA was generated by homologous recombination. After linearized with PacI restriction enzyme, vAd-PA was transfected into HEK-293 cell line by lipofection. Expression of PA in 293 cells was confirmed by SDS-PAGE and Western-blot. The BALB/c mice were intramuscularly inoculated with the recombinant adenovirus and the specific antibody was detected by ELISA . The induced antibody titer was 2.8×103 (Geometric Mean).

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    Construction of Recombinant Adenovirus Expressing Protective Antigen of Bacillus Anthracis and Analysis of Immuno-responses

    • 1. State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing 100071,China

    Abstract: In this study, we constructed recombinant adenovirus containing protective antigen gene of Bacillus anthracis. The PA gene was obtained from pcDNA3.1-PA vector by PCR , and cloned into plasmid pAdTrack-CMV. The positive clone was linearized with PmeI restriction enzyme and transformed into E. coli BJ5183 cells with the adenoviral plasmid pAdEasy-1. Then vAd-PA was generated by homologous recombination. After linearized with PacI restriction enzyme, vAd-PA was transfected into HEK-293 cell line by lipofection. Expression of PA in 293 cells was confirmed by SDS-PAGE and Western-blot. The BALB/c mice were intramuscularly inoculated with the recombinant adenovirus and the specific antibody was detected by ELISA . The induced antibody titer was 2.8×103 (Geometric Mean).

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