Table 1.

Vectors and recombinant clones used in this study

VectorRelevant characteristicsaReference or source
E. coli
 Plasmids
  pUC18pBR322-derivedE. coli vector; bla67
  pBR329E. coli general-purpose vector15
  pIJ2921pUC-derived E. coli vector with a modified polylinker;bla33
  pIJ2925pUC-derivedE. coli vector with a modified polylinker;bla33
  pSU19pACYC184-derived E. coli vector;cat3
  pET-19bpBR322-derived E. coli T7 polymerase expresion vector(Novagen, Madison, Wis.)
  pMV303KpnI-PstI fragment ofactVI-ORF1–ORFA (nt 1406–2632) cloned in pSU19;cat20
  pMF1013SalI(11.8–13.3) fragment cloned in pIJ2925; carries the wild-type actII-ORF4 gene;bla18
  pMF2001.2NaeI-SmaI fragment (nt 1914–2231) of the actVI intergenic region, cloned in pIJ2925 (HincII site);bla20
  pMF2065447-bpMboII fragment carrying theactIII-actI intergenic region, cloned in pIJ2925,bla19, 23
  pPAC2SphI-SacI fragment (positions 13.4–14.1) of the actIII-actI intergenic region, cloned in pIJ2925; bla19
  pPAC6Same as pMF1013; carries the actII-ORF4-177 gene from S. coelicolor JF1; blaThis work
  pPAC10NdeI-BglII fragment carrying the recombinant wild-type actII-ORF4 gene cloned in pET-19b (NdeI-BamHI sites); blaThis work
  pPAC14Same as pPAC10 but with the actII-ORF4-177 gene; blaThis work
 Phage
  M13mp18E. coli phage vector for DNA sequencing67
Streptomyces
 Plasmids
  pIJ486High-copy-number pIJ101 derivative vector,tsr63
  pIJ80Unstable SCP2*derivative plasmid; neo60a
  pIJ702High-copy-number vector; mel tsr34
  pIJ941Low-copy-number SCP2* derivative plasmid; tsr hyg40
  pMF1100SalI fragment (positions 11.8–13.3) cloned in pIJ941; carries the wild-type actII-ORF4 gene;hyg tsr18
  pMF1125SalI fragment (positions 11.8–13.3) cloned in pIJ702; carries the wild-type actII-ORF4 gene;tsrThis work
  pMF1123EcoRI-HindIII fragment of pMF2001.2 cloned in pIJ486; tsrThis work
  pMF1135EcoRI-HindIII fragment of pMF2065 cloned in pIJ486; tsrThis work
  pPAS1BglII fragment of pPAC6 cloned in pPAS3; carries the actII-ORF4-177 gene; hygThis work
  pPAS3pIJ941-derivd vector with a frameshit within the thiostrepton resistance gene; hyg44
  pPAS4SalI fragment (positions 11.8–13.3) cloned in pPAS3; carries the wild-type actII-ORF4 gene;hyg44
 Phages
  PM1φC31-derived vector; att tsr hyg41
  φAB9BglII fragment from pMF1013 cloned in the BamHI-BglII sites of PM1; hygThis work
  • a Abbreviations: bla, ampicillin resistance gene; cat, chloramphenicol acetyltransferase gene; hyg, hygromicin resistsance gene;neo, neomycin resistance gene; tsr, thiostrepton resistance gene; mel, tyrosinase gene; nt, nucleotides.