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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.

Please note: Wiley-Blackwell are not responsible for the content or functionality of any supporting materials supplied by the authors. Any queries (other than missing material) should be directed to the corresponding author for the article.

FilenameFormatSizeDescription
EMI_2227_sm_fS1.eps175KSupporting info item
EMI_2227_sm_fS2_1.eps648KSupporting info item
EMI_2227_sm_fS2_2.eps530KSupporting info item
EMI_2227_sm_fS2_3.eps567KSupporting info item
EMI_2227_sm_fS2_4.eps698KSupporting info item
EMI_2227_sm_fS2_5.eps521KSupporting info item
EMI_2227_sm_fS2_6.eps560KSupporting info item
EMI_2227_sm_fS2_7.eps631KSupporting info item
EMI_2227_sm_fS2_8.eps616KSupporting info item
EMI_2227_sm_fS2_9.eps524KSupporting info item
EMI_2227_sm_fS2_10.eps507KSupporting info item
EMI_2227_sm_fS3.eps227KSupporting info item
EMI_2227_sm_fS4.eps198KSupporting info item
EMI_2227_sm_tS1_1.eps503KSupporting info item
EMI_2227_sm_tS1_2.eps446KSupporting info item
EMI_2227_sm_tS1_3.eps440KSupporting info item
EMI_2227_sm_tS1_4.eps355KSupporting info item
EMI_2227_sm_tS2.eps397KSupporting info item

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.