File S1 - Function and Evolution of Two Forms of SecDF Homologs in Streptomyces coelicolor

Figures S1–S4, Tables S1 and S2. Figure S1. Identify the activities of secD-F and secDF gene promoters and gene expression profile. (A) Expression level of egfp in LM6 and ZJUZ39-41 by qPCR, the expression of egfp in LM1 was set as reference. (B) Detecting the expression of EGFP protein alone the time of cultivation by SDS-PAGE and Western blotting. WB: Western blotting; CB: Commassie Blue Staining. Figure S2. PCR and Southern blot verification of gene knock-out. (A) PCR analysis for disruption of secD-F genes. The primer pair secD_p_F vs secF_right_arm_R (Table S1) was used. (B) PCR analysis for disruption of secDF gene. The primer pair secDF_p_F vs secDF_right_arm_R (Table S1) was used. (C) Southern blot analysis for disruption of secD-F genes, digested by BamHI. (D) Southern blot analysis for disruption of secDF gene, digested by BamHI. Figure S3. Phenotypic analysis on morphogenesis between S. coelicolor wild type and mutants. (A) MM 60 h, (B) SMMS 84 h, (C) MSF 72 h, (D) R5 36 h. Figure S4. Semi-quantitative assay of extracellular AmlC activity. Strains ZJUZ27-30 were grown on MM media containing 0.2% soluble starch for 5 days before staining with Lugol’s solution. Table S1. Oligonucleotides used in this study. Table S2. Distribution of SecDF homologs in Streptomyces species and other species depicted in Figure 4. (PDF)