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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

Table 1

Genotypic characteristics of Escherichia coli and Bacillus anthracis strains and plasmids and primers used to determine avirulent B. anthracis strain with molecular assay targets

Strain, plasmid, primer
 Genotype
 Reference/source
Strain
DH5α F− Φ80lacZΔM15 Δ(lacZYA-argF) U169 recA1 endA1 hsdR17(rk−, mk+) phoA supE44 thi-1 gyrA96 relA1 λ− Laboratory collection
SCS110 rpsL thr leu endA thi-1 lacY galK galT ara tonA tsx dam dcm supE44 Δ(lac-proAB) Stratagene (La Jolla, CA, USA)
SM10 thi thr leu tonA lacY supE recA::RP4–2-Tc::Mu KmR λpir (9)
S17.1 hsdR pro recA, RP4–2 in chromosome, Km::Tn7 (Tc::Mu) (9)
DH5α/pSS1827 F− Φ80lacZΔM15 Δ(lacZYA-argF) U169 recA1 endA1 hsdR17(rk−, mk+) phoA supE44 thi-1 gyrA96 relA1 λ−, pSS1827 (Replicon fusion of pBR322 and pRK2013 at EcoRI and SalI sites) (10)
BA500 B. anthracis Sterne 34F2 Laboratory collection
BAP482 BA500 Δcya and Δlef (double toxin deletion) (11)
BAP417 BA500 Δcya, Δlef, and ΔpagA (triple toxin deletion) (11)
BAP708
 BA417 with construct 4 (Signatures 1–5)
 This study
Plasmid Description Source
pT7 Blue Cloning vector Novagen-MilliporeSigma, St. Louis, MO, USA
pT7 Blue::4 Construct 4 (Signatures 1–5) This study
pRP1091
 Δlef derivative of shuttle vector pRP1028
 (11)
Primer Sequence, 5′ → 3′ Application
RP411 TTTCACACAGGAAACAGCTATGACC Amplify constructs
RP645 CCAGTCACGACGTTGTAAAACGAC Amplify constructs
SS2178 GTAAATTATTTAGCAAGTAAATTTTGGTG Sequence constructs
RP214 TATGGTCTCGGATCCTTTGGCTTTAACGAAATGTATGTGC Diagnose, sequence lef
RP215 TATGGTCTCCGGCCGTTTCAGTTATTCATTCTGGATAGTC Diagnose, sequence lef
SS2164 CACGAGAAGAGTATTTAAAGAAAATC Diagnose lef
SS2165 AACTATAGGACAATATTCATTACCATG Diagnose lef
SS2166 ATATCAAGTTTAATTGTTAAGTTTGAAGG Diagnose cya
SS2167 CCCGCGGCCGCAACCAAATGGTTTTCATTTCTTAG Diagnose cya
SS2168 CGCATATAAGCAAATACTTAATTGGTC Diagnose pagA
SS2169 GGATAGGGTTTAACAACTTAATAATCCC Diagnose pagA

Main Article

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