Construction of the Genomic Expression Library of Bacillus anthracis for the Immunomic Analysis
2010
Park, M.K., Hanyang University, Ansan, Republic of Korea | Jung, K.H., Hanyang University, Ansan, Republic of Korea | Kim, Y.H., Center for Infectious Disease Research, National Institute of Health, Seoul, Republic of Korea | Rhie, G.E., Center for Infectious Disease Research, National Institute of Health, Seoul, Republic of Korea | Chai, Y.G., Hanyang University, Ansan, Republic of Korea | Yoon, J.W., Hanyang University, Ansan, Republic of Korea
As the causative agent of Anthrax, Bacillus anthracis causes an acute fatal disease in herbivores such as cattle, sheep, and horses as well as humans. The therapeutics and prevention of anthrax currently available are based on antibiotics and the live attenuated vaccine strains, which may be problematic due to the emergency of antibiotic resistant strains or residual virulence in those vaccine strains. Therefore, it has been required to develop novel therapeutics and vaccines which are safer and applicable to humans. Recently, the development of the multivalent vaccine targeting both spores and vegetative cells of B. anthracis along with anthrax toxin has been reported. In our attempts to screen potential candidates for those multivalent vaccines, the whole genomic expression library of B. anthracis was constructed in this study. To the end, the partial digests of the genomic DNA from B. anthracis (ATCC 14578) with Sau3AI were ligated with the inducible pET30abc expression vectors, resulting in approximately 1×10∨5 clones in E. coli BL21(DE3). The redundancy test by DNA nucleotide sequencing was performed for the randomly selected 111 clones and found 56 (50.5%) B. anthracis genes, 17 (15.3%) vector sequences, and 38 (34.2%) unknown genes with no sequence homology by BLAST. An inducible expression of the recombinant proteins was confirmed by Western blot. Interestingly, some clones could react with the antiserum against B. anthracis. These results imply that the whole genomic library constructed in this study can be applied for analyzing the immunomes of B. anthracis.
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