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