Additional file 1 of Effect of the TetR family transcriptional regulator Sp1418 on the global metabolic network of Saccharopolyspora pogona
Creators: name: He, Haocheng
name: Yuan, Shuangqin
name: Hu, Jinjuan
name: Chen, Jianming
name: Rang, Jie
name: Tang, Jianli
name: Liu, Zhudong
name: Xia, Ziyuan
name: Ding, Xuezhi
name: Hu, Shengbiao
name: Xia, Liqiu
Publisher: name: figshare
Year: 2020
Subjects: subject: Biophysics
subject: Biochemistry
subject: Microbiology
subject: FOS: Biological sciences
subject: Cell Biology
subject: Genetics
subject: Molecular Biology
subject: Biotechnology
subject: Biological Sciences not elsewhere classified
subject: Developmental Biology
subject: Infectious Diseases
subject: FOS: Health sciences
subject: Plant Biology

description: Additional file 1: Figure S1. Mass spectrum identification of butenyl-spinosyns. MS identification results showed that MS parent ion [M+H]+ = 633 (black arrow) contained 617 (M+H+) (m/z) ion data and a rhamnose ion fragment of 189 molecular mass, which was confirmed as a butenyl-spinosyn component. Figure S2. The sporulation phenotypes in wild type, S. pogona-Δsp1418 and S. pogona-Sp1418. S. pogona-Sp1418 did not produce spores or was almost invisible, and the amount of spores of S. pogona-Δsp1418 is much more than that of S. pogona, but there was no significant difference in the spore morphology of S. pogona and S. pogona-Δsp1418. Figure S3. Tricine-SDS-PAGE analysis of heterologously expressed protein Sp1418. Coomassie Brilliant Blue staining of Tricine-SDS-PAGE showing heterologous protein Sp1418 expressed in the supernatants of E. coli BL21 bearing recombinant plasmid after IPTG induction and ultrasonication. M: 66 kDa protein marker; 1: Samples from E. coli BL21 as a negative control 2: Samples from E. coli BL21 contained the recombinant plasmid. Figure S4. SDS-PAGE gel analysis of total proteins. M: Protein marker; 1: Samples from 96 h S. pogona; 2: Samples from 96 h S. pogona-Sp1418 cells; 3: Samples from 96 h S. pogona-Δsp1418 cells. Compared with the three strains, there are many distinctive bands, and totally 9 proteins were identified via 1D-LC–MS/MS. Figure S5. Construction of pKCcas9d-sgRNA-UHA-DHA and pOJ260-PermE-sp1418. A. Construction of plasmid pKCcas9d-sgRNA-UHA-DHA; B. Construction of plasmid pOJ260-PermE-sp1418.Figure S6. Recombination schematic diagram of vetor pOJ260-PermE-sp1418 and pKCcas9d-sgRNA-UHA-DHA. A. Recombination schematic diagram of vetor pOJ260-PermE-sp1418; B. Recombination schematic diagram of vetor pKCcas9d-sgRNA-UHA-DHA. Figure S7. Identification of S. pogona-Δsp1418 and S. pogona-Sp1418. A: Identification of PermE-sp1418 fragment in S. pogona and S. pogona-Sp1418. M: DL 2000 DNA marker; 1: PCR products of S. pogona with primer pair perm-F/tetR-R; 2: PCR products of S. pogona-Sp1418 with primer pair perm-F/tetR-R; B: PCR amplification of aac(3)IV gene in S. pogona and S. pogona-Sp1418. M: DL 2000 DNA marker; 1–2: PCR products of S. pogona with primer pair Apr-F/Apr-R; 3: PCR products of S. pogona-Sp1418 with primer pair Apr-F/Apr-R; C: Identification of aac(3)IV gene in S. pogona and S. pogona-Δsp1418. M: DL 2000 DNA marker; 1: PCR products of S. pogona with primers Apr-F/Apr-R; 2: PCR products of S. pogona-Δsp1418 with primers Apr-F/Apr-R; D: Identification of sp1418 gene in S. pogona and S. pogona-Δsp1418 M: DL 2000 DNA marker; 1: PCR products of S. pogona with primers tetR-P-F/tetR-P-R; 2: PCR products of S. pogona-Δsp1418 with primers tetR-P-F/tetR-P-R.