Summary
The availability of the Bacillus anthracis genome sequence allowed for in silico selection of a few hundred open reading frames (ORFs) as putative vaccine candidates. To screen such a vast number of candidate ORFs, without resorting to laborious cloning and protein purification procedures, methods were developed for generation of PCR elements, compatible with in vitro transcription-translation and immunoprecipitation, as well as with their evaluation as DNA vaccines. Protocols will be provided for application of these methods to analyze the anti-B. anthracis antibody repertoire of hyperimmune sera or sera from convalescent and from DNA-vaccinated animals.
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References
Mock, M. and Fouet, A. (2001) Anthrax. Annu. Rev. Microbiol. 55, 647–671.
Friedlander, A. M., Welkos, S. L., and Ivins, B. E. (2002) Anthrax vaccines. Curr. Top. Microbiol. Immunol. 271, 33–60.
Adu-Bobie, J., Capecchi, B., Serruto, D., Rappuoli, R., and Pizza, M. (2002) Two years into reverse vaccinology. Vaccine 21, 605–610.
Read, T. D., Peterson, S. N., Tourasse, N., et al. (2003) The genome sequence of Bacillus anthracis Ames and comparison to closely related bacteria. Nature 423, 81–86.
Okinaka, R. T., Cloud, K., Hampton, O., et al. (1999) Sequence and organization of pXO1, the large Bacillus anthracis plasmid harboring the anthrax toxin genes. J. Bacteriol. 181, 6509–6515.
Ariel, N., Zvi, A., Makarova, K. S., et al. (2003) Genome-based bioinformatic selection of chromosomal Bacillus anthracis putative vaccine candidates coupled with proteomic identification of surface-associated antigens. Infect. Immun. 71, 4563–4579.
Ariel, N., Zvi, A., Grosfeld, H., et al. (2002) Search for potential vaccine candidate open reading frames in the Bacillus anthracis virulence plasmid pXO1: in silico and in vitro screening. Infect. Immun. 70, 6817–6827.
Johnson, J. L. (1994) Similarity analysis of DNAs, in Methods for General and Molecular Bacteriology, (Gerhardt, P., Murray, R. G. E., Wood, W. A., and Krieg, N. R., eds.), ASM, Washington, DC, pp. 655–682.
Chitlaru, T., Ariel, N., Zvi, A., et al. (2004) Identification of chromosomally encoded membranal polypeptides of Bacillus anthracis by a proteomic analysis: Prevalence of proteins containing S-layer homology domains. Proteomics 4, 677–691.
Altboum, Z., Gozes, Y., Barnea, A., Pass, A., White, M., and Kobiler, D. (2002) Postexposure prophylaxis against anthrax: evaluation of various treatment regimens in intranasally infected guinea pigs. Infect. Immun. 70, 6231–6241.
Grosfeld, H., Cohen, S., Bino, T., et al. (2003) Effective protective immunity to Yersinia pestis infection conferred by DNA vaccine coding for derivatives of the F1 capsular antigen. Infect. Immun. 71, 374–383.
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© 2007 Humana Press Inc., Totowa, NJ
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Gat, O., Grosfeld, H., Shafferman, A. (2007). In Vitro Screen of Bioinformatically Selected Bacillus anthracis Vaccine Candidates by Coupled Transcription, Translation, and Immunoprecipitation Analysis. In: Grandi, G. (eds) In Vitro Transcription and Translation Protocols. Methods in Molecular Biology™, vol 375. Humana Press. https://doi.org/10.1007/978-1-59745-388-2_11
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DOI: https://doi.org/10.1007/978-1-59745-388-2_11
Publisher Name: Humana Press
Print ISBN: 978-1-58829-558-3
Online ISBN: 978-1-59745-388-2
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