ArticlePloS one2015
Improvements to a Markerless Allelic Exchange System for Bacillus anthracis.
Article in PloS one, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 23 papers.
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Who cites it
23 citing papers in PubMed.
- Functional disparities of malonyl-ACP decarboxylase betweenApplied and environmental microbiology · 2025Article
- Pleiotropic functions of CpdB in Bacillus anthracis.World journal of microbiology & biotechnology · 2025Article
- EsxA, a type VII secretion system-dependent effector, reveals a novel function in the sporulation of Bacillus cereus ATCC14579.BMC microbiology · 2024Article
- YfmR is a translation factor that prevents ribosome stalling and cell death in the absence of EF-P.Proceedings of the National Academy of Sciences of the United States of America · 2024Article
- YfmR is a translation factor that prevents ribosome stalling and cell death in the absence of EF-P.bioRxiv : the preprint server for biology · 2023Article
- Characterization of Lactobacilli Phage Endolysins and Their Functional Domains-Potential Live Biotherapeutic Testing Reagents.Viruses · 2023Article
- Beyond the spore, the exosporium sugar anthrose impacts vegetative Bacillus anthracis gene regulation in cis and trans.Scientific reports · 2023Article
- Role of novel polysaccharide layers in assembly of the exosporium, the outermost protein layer of the Bacillus anthracis spore.Molecular microbiology · 2022Article
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- Silkworm model forVirulence · 2021Article
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- barCoder: a tool to generate unique, orthogonal genetic tags for qPCR detection.BMC bioinformatics · 2021Article
- Convergent evolution of diverse Bacillus anthracis outbreak strains toward altered surface oligosaccharides that modulate anthrax pathogenesis.PLoS biology · 2020Article
- Increased Excess Intracellular Cyclic di-AMP Levels Impair Growth and Virulence of Bacillus anthracis.Journal of bacteriology · 2020Article
- Linking Geospatial and Laboratory Sciences to Define Mechanisms behind Landscape Level Drivers of Anthrax Outbreaks.International journal of environmental research and public health · 2019Article
- TheMicrobiology spectrum · 2019Review
- Highly Efficient Genome Engineering inFrontiers in microbiology · 2019Article
- Article
- A Standard Method To Inactivate Bacillus anthracis Spores to Sterility via Gamma Irradiation.Applied and environmental microbiology · 2018Article
- Phage Therapy: What Have We Learned?Viruses · 2018Review
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Authors and funding
2 authors.
Funding
Abstract
A system was previously developed for conducting I-SceI-mediated allelic exchange in Bacillus anthracis. In this system, recombinational loss of a chromosomally-integrated allelic exchange vector is stimulated by creation of a double-stranded break within the vector by the homing endonuclease I-SceI. Although this system is reasonably efficient and represents an improvement in the tools available for allelic exchange in B. anthracis, researchers are nonetheless required to "pick and patch" colonies in order to identify candidate "exchangeants." In the present study, a number of improvements have been made to this system: 1) an improved I-SceI-producing plasmid includes oriT so that both plasmids can now be introduced by conjugation, thus avoiding the need for preparing electro-competent cells of each integration intermediate; 2) antibiotic markers have been changed to allow the use of the system in select agent strains; and 3) both plasmids have been marked with fluorescent proteins, allowing the visualization of plasmid segregation on a plate and obviating the need for "picking and patching." These modifications have made the process easier, faster, and more efficient, allowing for parallel construction of larger numbers of mutant strains. Using this improved system, the genes encoding the tripartite anthrax toxin were deleted singly and in combination from plasmid pXO1 of Sterne strain 34F2. In the course of this study, we determined that DNA transfer to B. anthracis could be accomplished by conjugation directly from a methylation-competent E. coli strain.
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