Burkholderia cenocepacia conditional growth mutant library created by random promoter replacement of essential genes
Article first published online: 7 FEB 2013
© 2013 The Authors. MicrobiologyOpen published by Blackwell Publishing Ltd.
This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
Volume 2, Issue 2, pages 243–258, April 2013
How to Cite
Bloodworth, R. A. M., Gislason, A. S. and Cardona, S. T. (2013), Burkholderia cenocepacia conditional growth mutant library created by random promoter replacement of essential genes. MicrobiologyOpen, 2: 243–258. doi: 10.1002/mbo3.71
- Issue published online: 8 APR 2013
- Article first published online: 7 FEB 2013
- Manuscript Accepted: 8 JAN 2013
- Manuscript Revised: 24 DEC 2012
- Manuscript Received: 24 OCT 2012
- Manitoba Health and Research Council
- Canadian Institutes of Health Research
- Natural Sciences and Engineering Research Council
- Paul Thorlakson Foundation Fund
- University of Manitoba
Figure S1. The number of essential genes as a function of genome size. The overall number of essential genes in 14 bacterial genomes is plotted against the total number of genes in the same genome. Data on gene essentiality for all genomes with the exception of data noted with asterisks were collected from Gerdes et al. (2006); gene essentiality of Caulobacter crescentus, Mycobacterium tuberculosis, and Salmonella typhi were obtained from Christen et al. (2011), Griffin et al. (2011), and Langridge et al. (2009), respectively. Mutants were generated by random or targeted transposon mutagenesis, and mutants were propagated. Mutant lack of survival was considered criteria for defining the interrupted gene as essential. Gray and white squares indicate that the data were obtained from propagating the mutants within a population or clonally, respectively. Mge, Mycoplasma genitalium; Hin, Haemophilus influenzae Rd; Hpy, Helicobacter pylori G27; Sau, Staphylococcus aureus RN4220; Mtu, M. tuberculosis H37Rv; Eco, Escherichia coli; Sty, S. typhi; Ccr, C. crescentus; Bsu, Bacillus subtilis; PAO1, Pseudomonas aeruginosa PAO1; PA14, P. aeruginosa PA14.
Figure S2. Transposon insertions relative to the start of relevant coding sequence. (A) Histogram of the distance from insertions into putative intergenic regions to the putative start codon of the downstream gene measured in base pairs. (B) Histogram of the distance from insertions inside of putative coding sequences to the start codon of the surrounding gene measured as a percentage of total gene length.
Figure S3. Conditional growth mutants show selective hypersensitivity. Mutants were grown in rhamnose concentration gradients estimated to produce more than 30% of wild-type growth and challenged with either novobiocin or chloramphenicol at the IC30 of the wild-type. Circles represent CG mutants of the direct target of novobiocin, gyrB. Green crosses represent a CG mutant of the electron transfer flavoprotein gene (etfA). Black crosses correspond to nonsensitive mutants. Mutants of gyrB show hypersensitivity to novobiocin when grown in rhamnose concentrations producing 30–60% of wild-type growth but not when grown in rhamnose concentrations producing 80–100% of wild-type growth. A CG mutant of etfA shows intermediate hypersensitivity. None of the mutants showed hypersensitivity to chloramphenicol. Error bars represent 1 standard deviation calculated from two biological replicates.
Table S1. Mutants found in this study.
Table S2. Primers.
Table S3. Genes found in this study with essential orthologs in Escherichia coli or Pseudomonas aeruginosa.
Table S4. Burkholderia species-specific putative essential operons.
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