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

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

By: Orit Gat², Haim Grosfeld², Avigdor Shafferman²

Abstract

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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.

Affiliation(s): (2) Department of Biochemistry and Molecular Genetics, Israel Institute for Biological Research, Ness-Ziona, Israel

Book Title: In Vitro Transcription and Translation Protocols

Series: Methods in Molecular Biology | Volume: 375 | Pub. Date: May-03-2007 | Page Range: 211-233 | DOI: 10.1007/978-1-59745-388-2_11

Subject: Genetics/Genomics

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

References

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