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Hierarchical control of virginiamycin production in Streptomyces virginiae by three pathway-specific regulators: VmsS, VmsT and VmsR, by N. Pulsawat, S. Kitani, E. Fukushima and T. Nihira

Microbiology vol. 155, part 4, pp. 1250 - 1259

Table S1. Oligonucleotides used in this study [PDF] (45 kb)

Fig. S1. Disruption of vmsS, vmsT and vmsR genes in S. virginiae. (a-c) Schematic representation of the strategy for the gene disruption of vmsS (a), vmsT (b) and vmsR (c). (d) Southern blot hybridization analysis of the wild-type strain, and the mutants IC102, IC103 and IC104. Genomic DNAs were digested with BamHI, SacI and SacI for vmsS, vmsT and vmsR mutants, respectively. Plasmid-integrated strains are mutants in which the disruption plasmids were introduced into the genome by single-crossover homologous recombination. [PDF] (69 kb)

Fig. S2. Transcriptional analysis of vms genes in the corresponding mutant. Transcripts from mutants (a) IC102 (ΔvmsS), (b) IC103 (ΔvmsT) and (c) IC104 (ΔvmsR) were analysed. Total RNAs were extracted from mycelia harvested at 14 h cultivation. Each transcript (I) was amplified with the primer sets as used in Fig. 2(a) of the main paper, and each transcript (II) was amplified with primer sets that were designed (Table S1) to detect cDNA corresponding to downstream of the mutated region. The varR gene was used as a control. -RT indicates PCR product performed without the initial reverse transcription step in RT-PCR. The cycle numbers for PCR were 27 cycles for vms genes and 25 cycles for varR. [PDF] (108 kb)

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