In vitro screen of bioinformatically selected Bacillus anthracis vaccine candidates by coupled transcription, translation, and immunoprecipitation analysis

Methods Mol Biol. 2007:375:211-33. doi: 10.1007/978-1-59745-388-2_11.

Abstract

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.

Publication types

  • Review

MeSH terms

  • Animals
  • Anthrax / immunology
  • Anthrax / microbiology*
  • Anthrax / prevention & control
  • Anthrax Vaccines / genetics*
  • Anthrax Vaccines / immunology*
  • Antigens, Bacterial / genetics
  • Antigens, Bacterial / immunology
  • Antigens, Bacterial / physiology
  • Bacillus anthracis / genetics
  • Bacillus anthracis / immunology*
  • Computational Biology
  • Electroporation
  • Genome, Bacterial / immunology
  • Immunoprecipitation*
  • In Vitro Techniques
  • Open Reading Frames
  • Protein Biosynthesis*
  • Transcription, Genetic*

Substances

  • Anthrax Vaccines
  • Antigens, Bacterial