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An attenuated strain of Bacillus anthracis (CDC 684) has a large chromosomal inversion and altered growth kinetics

Richard T Okinaka12*, Erin P Price1, Spenser R Wolken1, Jeffrey M Gruendike1, Wai Kwan Chung1, Talima Pearson1, Gary Xie2, Chris Munk2, Karen K Hill2, Jean Challacombe2, Bruce E Ivins3, James M Schupp4, Stephen M Beckstrom-Sternberg14, Arthur Friedlander3 and Paul Keim124

Author Affiliations

1 Center for Microbial Genetics and Genomics, Northern Arizona University, Flagstaff, AZ 86011, USA

2 Bioscience Division, Los Alamos National Laboratory, Los Alamos, NM 87545, USA

3 Bacteriology Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, MD 21702-5011, USA

4 Pathogen Genomics Division, Translational Genomics Research Institute, Phoenix, AZ 85004, USA

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BMC Genomics 2011, 12:477  doi:10.1186/1471-2164-12-477

Published: 30 September 2011

Additional files

Additional file 1:

Figure S1: MAMA real time PCR. Example of MAMA Real Time PCR to Differentiate between a 3.3 Mbp inverted configuration in CDC 684 and the wild type configuration in Bacillus anthracis. The primers depicted in this figure were designed to detect the left configuration for the wild-type and/or the inverted sequence for the CDC 684 genome. The fixed common primer, CP Left-inv-F was paired with both Right-inv-F and the Left inv-R primers. While the wild type Ames Ancestor can be amplified by the two LEFT inversion primers (Left inv F and Left inv R); the left inversion site in CDC 684 can only be amplified by the CP left-inv-F and Right inv-F because the Right inversion forward site normally situated 3.3 Mbp distal, has relocated to within 450 bp of the CP left-inv-F primer. Note that these two panels represent samples with the same three primers and that the positive allele will amplify orders of magnitudes faster than the negative allele. The delta CT (cross-over threshold) values are the differences between a specific linear portion of the target sequence (in cycles) versus the number of cycles to obtain the same (threshold) signal in the non-target sequence (e.g....the delta CT for CDC 684 vs Ames is 13.4.

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Additional file 2:

Table S1: The SYBR MAMA primer sets for 10 Vollum lineage canSNP Assays.

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