In vitro screen of bioinformatically selected Bacillus anthracis vaccine candidates by coupled transcription, translation, and immunoprecipitation analysis.
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.
Gat
Orit
O
Department of Biochemistry and Molecular Genetics, Israel Institute for Biological Research, Ness Ziona, Israel.
Grosfeld
Haim
H
Shafferman
Avigdor
A
D000818:Animals; D000881:Anthrax / Q000276:immunology / Q000382:microbiology* / Q000517:prevention & control; D022122:Anthrax Vaccines / Q000235:genetics* / Q000276:immunology*; D000942:Antigens, Bacterial / Q000235:genetics / Q000276:immunology / Q000502:physiology; D001408:Bacillus anthracis / Q000235:genetics / Q000276:immunology*; D019295:Computational Biology; D018274:Electroporation; D016680:Genome, Bacterial / Q000276:immunology; D047468:Immunoprecipitation; D066298:In Vitro Techniques; D016366:Open Reading Frames; D014176:Protein Biosynthesis; D014158:Transcription, Genetic
D022122:Anthrax Vaccines; D000942:Antigens, Bacterial