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Volume 24, Number 4—April 2018
Research

Avirulent Bacillus anthracis Strain with Molecular Assay Targets as Surrogate for Irradiation-Inactivated Virulent Spores

Roger D. Plaut, Andrea B. Staab, Mark A. Munson, Joan S. Gebhardt, Christopher P. Klimko, Avery V. Quirk, Christopher K. Cote, Tony L. Buhr, Rebecca D. Rossmaier, Robert C. Bernhards, Courtney E. Love, Kimberly L. Berk, Teresa G. Abshire, David A. Rozak, Linda C. Beck, Scott Stibitz, Bruce G. Goodwin, Michael A. Smith, and Shanmuga SozhamannanComments to Author 
Author affiliations: Food and Drug Administration, Silver Spring, Maryland, USA (R.D. Plaut, S. Stibitz); Naval Surface Warfare Center, Dahlgren, Virginia, USA (A.B. Staab, T.L. Buhr, L.C. Beck); Naval Medical Research Center, Fort Detrick, Maryland, USA (M.A. Munson, J.S. Gebhardt); US Army Medical Research Institute of Infectious Diseases, Fort Detrick (C.P. Klimko, A.V. Quirk, C.K. Cote, T.G. Abshire, D.A. Rozak); US Army Edgewood Chemical and Biological Center, Aberdeen Proving Ground, Maryland, USA (R.D. Rossmaier, R.C. Bernhards, C.E. Love, K.L. Berk); Defense Threat Reduction Agency, Fort Belvoir, Virginia, USA (R.C. Bernhards); Joint Research and Development, Inc., Stafford, Virginia, USA (C.E. Love, L.C. Beck); Defense Biological Product Assurance Office, Frederick, Maryland, USA (B.G. Goodwin, M.A. Smith, S. Sozhamannan); The Tauri Group, Inc., Alexandria, Virginia, USA (S. Sozhamannan)

Main Article

Figure 1

Verification of toxin gene deletions and the genetic structure of the construct 4 cassette in Bacillus anthracis surrogate strain. A) PCR verification of toxin gene deletions in BA500 (Sterne 34F2) derivatives. Single colonies were processed and used as templates for PCR with respective primers as described in Methods. For each strain, primers were used to amplify, from left to right, the regions of cya (SS2166/SS2167), lef (SS2164/SS2165), and pagA (SS2168/SS2169) on pXO1. B) Schematic represen

Figure 1. Verification of toxin gene deletions and the genetic structure of the construct 4 cassette in Bacillus anthracis surrogate strain. A) PCR verification of toxin gene deletions in BA500 (Sterne 34F2) derivatives. Single colonies were processed and used as templates for PCR with respective primers as described in Methods. For each strain, primers were used to amplify, from left to right, the regions of cya (SS2166/SS2167), lef (SS2164/SS2165), and pagA (SS2168/SS2169) on pXO1. B) Schematic representation of BAP708 (construct 4) cassette. Green bars represent the PCR signatures, red bars represent bar codes, and black boxes represent stop codons in all 3 open reading frames. XbaI sites at the ends of the cassette used in subcloning of the insert are marked. C) PCR verification of the presence of construct 4 synthetic sequence cassette in BAP708, using primers immediately flanking the lef deletion region (RP214 and RP215). Strains and PCR primers are listed in Table 1. Ladder indicates size in kbps. WT, wild-type.

Main Article

Page created: March 19, 2018
Page updated: March 19, 2018
Page reviewed: March 19, 2018
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