Transposon mutagenesis of Bacillus anthracis strain Sterne using Bursa aurealis

Plasmid
Christina TamDominique M Missiakas

Abstract

Bacillus anthracis, a spore forming Gram-positive microbe, is the causative agent of anthrax. Although plasmid encoded factors such as lethal toxin (LeTx), edema toxin (EdTx), and gamma-poly-d-glutamic acid (PGA) capsule are known to be required for disease pathogenesis, B. anthracis genes that enable spore invasion, phagosomal escape and macrophage replication are still unknown. To establish transposon mutagenesis as a tool for the characterization of anthrax genes, we employed the mariner-based mini-transposon Bursa aurealis in B. anthracis strain Sterne 7702. B. aurealis carrying an erythromycin resistance cassette and its cognate transposase were delivered by transformation of two plasmids. B. aurealis transposition can be selected for by temperature shift to prevent plasmid replication and by screening colonies for erythromycin resistance. Using inverse polymerase chain reaction, DNA fragments of 129 random erythromycin-resistant transposon mutants were amplified and submitted to DNA sequence analysis. These studies demonstrate that B. aurealis inserts randomly into the genome of B. anthracis and can therefore be employed for finding genes involved in virulence.

References

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Aug 12, 2004·Proceedings of the National Academy of Sciences of the United States of America·Taeok BaeDominique M Missiakas

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Citations

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Apr 15, 2008·Applied and Environmental Microbiology·Christophe BordiJohn D Helmann
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May 23, 2012·Journal of Bacteriology·Sao-Mai Nguyen-MauOlaf Schneewind
Jun 12, 2012·Journal of Bacteriology·Gabriella GarufiDominique Missiakas
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Nov 12, 2014·Journal of Bacteriology·Megan Liszewski ZillaDominique Missiakas
Mar 18, 2015·Journal of Bacteriology·So-Young OhOlaf Schneewind
Jul 29, 2015·Journal of Bacteriology·Sao-Mai Nguyen-MauOlaf Schneewind
Sep 24, 2015·Journal of Bacteriology·Megan Liszewski ZillaDominique Missiakas
Feb 11, 2014·Current Protocols in Microbiology·Olaf Schneewind, Dominique Missiakas
May 22, 2019·Microbiology Spectrum·Monika Ehling-SchulzTheresa M Koehler

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