In Vitro Screen of Bioinformatically Selected Bacillus anthracis Vaccine Candidates by Coupled Transcription, Translation, and Immunoprecipitation Analysis

  • Orit Gat
  • Haim Grosfeld
  • Avigdor Shafferman
Part of the Methods in Molecular Biology™ book series (MIMB, volume 375)


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.

Key Words

In vitro transcription in vitro translation reverse vaccinology vaccine development Bacillus anthracis bioinformatics geomics DNA vaccine gene gun immunoprecipitation serological analysis immunogenicity screen antigenicity screen 


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Copyright information

© Humana Press Inc., Totowa, NJ 2007

Authors and Affiliations

  • Orit Gat
    • 1
  • Haim Grosfeld
    • 1
  • Avigdor Shafferman
    • 1
  1. 1.Department of Biochemistry and Molecular GeneticsIsrael Institute for Biological ResearchNess-ZionaIsrael

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