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Table 1 Plasmids and bacterial strains used in this study

From: Metabolic engineering of roseoflavin-overproducing microorganisms

Plasmid or strain Relevant feature(s)a Source or references
Plasmids
 pMK-RQ-rosB-rosA-RFK Plasmid containing synthetic (codon-adapted) genes of rosB, rosA and RFK (sequence see Additional file 1: Figure S1); oriEc; KanR Life Technologies™ (California, USA)
 pHT01 Expression vector for intracellular production of recombinant proteins in Bacillus subtilis. The Pgrac promoter region consists of the groE promoter, the lacO operator and SDgsiB. Gene expression is stimulated by addition of IPTG; oriEc; AmpR; CmR MoBiTec GmbH (Göttingen, Germany)
 pRML1 pHT01 derivative containing rosB, rosA and RFK This study
 pGP888 Expression vector for intracellular production of recombinant proteins in Bacillus subtilis; the linearized plasmid integrates into chromosomal ganA; gene expression is stimulated by addition of xylose; PxylA; KanR [44]
 pRML2 pGP888 derivative containing rosB, rosA and RFK This study
 pMKEx2 Expression vector for intracellular production of recombinant proteins in Escherichia coli and Corynebacterium glutamicum (shuttle vector). Gene expression is stimulated by addition of IPTG; PT7, oriEc; oriCg; KanR [45]
 pRML3 pMKEx2 derivative containing rosB, rosA and RFK This study
 pHT01_ribMopt pHT01 derivative containing the riboflavin/roseoflavin transporter gene ribM; oriEc; AmpR; CmR [13]
 pRML4 pMKEx2 derivative containing rosB, rosA, RFK and ribM This study
 pSET152 Vector for conjugal transfer of DNA from E. coli to Streptomyces; disrupts attB site of Streptomyces davaonensis genome; non-replicative in Streptomyces; oriEc; ApmR [46]
 pSETPermErosArosB pSET152 derivative containing rosA* and rosB* under control of the promoter PermE* [17]
Strains
 E. coli
  Top10 For cloning work Invitrogen™ (California, USA)
  EC01 E. coli Top10 containing pHT01; AmpR This study
  EC02 E. coli Top10 containing pRML1; AmpR This study
  EC03 E. coli Top10 containing pHT01_ribMopt; AmpR This study
  EC04 E. coli Top10 containing pGP888; AmpR This study
  EC05 E. coli Top10 containing pRML2; AmpR This study
  EC06 E. coli Top10 containing pMKEx2; KanR This study
  EC07 E. coli Top10 containing pRML3; KanR This study
  EC08 E. coli Top10 containing pRML4; KanR This study
  GM2163/pUB306 Methylation deficient strain used for conjugation with S. davaonensis; StrR; CmR [47]
  EC09 E. coli GM2163/pUB306 containing pSETPermErosArosB; StrR; CmR This study
 B. subtilis
  WT1 Wild-type strain #1; B. subtilis riboflavin-overproducing strain (ROP aka. RÜP or BSHP) [13]
  C1 B. subtilis ROP containing pHT01 (empty plasmid); control strain; CmR This study
  RML1 B. subtilis ROP containing pRML1; IPTG-inducible; CmR This study
  C2 B. subtilis ROP containing pGP888 (empty plasmid); control strain; KanR This study
  RML2 B. subtilis ROP containing pRML2; xylose-inducible; KanR This study
  WT2 Wild-type strain #2; B. subtilis 168 strain Bacillus Genetic Stock Center (BGSC)
  C3 B. subtilis 168 containing pHT01 (empty plasmid); control strain; CmR This study
  RML3 B. subtilis 168 containing pRML1; IPTG-inducible; CmR This study
  C4 B. subtilis 168 containing pGP888 (empty plasmid); control strain; KanR This study
  RML4 B. subtilis 168 containing pRML2; xylose-inducible; KanR This study
 C. glutamicum
  WT Wild-type strain; C. glutamicum MB001(DE3) strain [45]
  C5–6 C. glutamicum MB001(DE3) containing pMKEx2 (empty plasmid); control strain; KanR This study
  RML5 C. glutamicum MB001(DE3) containing pRML3; IPTG-inducible; KanR This study
  RML6 C. glutamicum MB001(DE3) containing pRML4; IPTG-inducible; KanR This study
 S. davaonensis
  WT Wild-type strain; S. davaonensis JCM 4913 strain Japan Collection of Microorganisms
  C7 S. davaonensis containing pSET152 (empty plasmid); control strain; AmpR; ApmR This study
  RML7 S. davaonensis containing pSETPermErosArosB; AmpR; ApmR This study
  1. aP: promoter; SD: ribosomal binding site; ori: origin of replication; AmpR: ampicillin resistance; ApmR: apramycin resistance; CmR: chloramphenicol resistance; KanR: kanamycin resistance; rosB, rosA, RFK and ribM: codon optimized genes for expression in B. subtilis; rosA* and rosB*: native genes from S. davaonensis