Identification of Burkholderia multivorans ATCC 17616 genes induced in soil environment by in vivo expression technology
Article first published online: 19 APR 2010
© 2010 Society for Applied Microbiology and Blackwell Publishing Ltd
Volume 12, Issue 9, pages 2539–2558, September 2010
How to Cite
Nishiyama, E., Ohtsubo, Y., Nagata, Y. and Tsuda, M. (2010), Identification of Burkholderia multivorans ATCC 17616 genes induced in soil environment by in vivo expression technology. Environmental Microbiology, 12: 2539–2558. doi: 10.1111/j.1462-2920.2010.02227.x
- Issue published online: 3 SEP 2010
- Article first published online: 19 APR 2010
- Received 23 January, 2010; accepted 22 February, 2010.
Fig. S1. Determination of incubation period in soil for IVET screening. Approximately equal numbers of ATCC 17616ΔdapB and its derivative (an ATCC 17616ΔdapB derivative having a genomic insert of pEN3 whose dapB-lacZ cassette was expressed constitutively both in laboratory medium and soil) cells were mixed. The sterilized soil was inoculated with the mixture, and after appropriate periods, the cells recovered from the soil were plated on M9 agar containing X-gal, DAP and lysine. Results are averages of three independent experiments, and standard deviations are shown.
Fig. S2. Genomic context of mls in B. multivorans. For each mls, the DNA fragments cloned in pEN3 are indicated by arrows, and the arrowheads represent the genomic positions located just in front of dapB-lacZ cassette. The names of mls fusion clones are indicated beside the arrows, and the mls fusion clones used in the present analysis are depicted by red colour. The mls described in text is underlined. Grey pentagon indicates the putatively soil-induced genes, and the number in pentagon represents the locus tag with the prefix of BMULJ_. Black symbols represent genomic features, which were given the ‘repeat_region’ feature key in the GenBank-formatted annotation.
Fig. S3. Transcriptional extension reaching to dapB-lacZ cassette inserted at far downstream region. The pEN3 portion blocks the opposing transcription, thus allowing the RNA polymerase to reach the dapB-lacZ genes. This situation can be exemplified in an mls224 fusion clone.
Fig. S4. Exceptional gene disruption by integration of pEN3 derivative. The integration of a pEN3 derivative that carries an internal portion of a gene results in the disruption of the gene. This situation can be exemplified in an mls207 fusion clone, in which the plasmid integration site is within a gene coding for the protein with 2505 amino acid residues.
Table S1. List of mls that were identified only once and have not been confirmed for in vivo induction.
Table S2. Oligonucleotide primers used in this studya.
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Please note: Wiley Blackwell is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.