TABLE 1.

Plasmids

PlasmidRelevant characteristicsaReference(s) or source
pSK+Apr, lac promoter, pMB1 repliconStratagene
pSTV28Cmlr, lac promoter, p15A repliconTakara
pET100/DApr, T7 promoter, pMB1 repliconInvitrogen
pET101/DApr, T7 promoter, pMB1 repliconInvitrogen
pRD1pWE15::32,000-bp Sau3AI fragment containing the dha regulon (dhaBCDEFGHIKRT genes) of C. freundii 15, 16, 41
pFL1pWE15::13,500-bp Sau3AI fragment containing dhaBCEFGHIT genes of Clostridium pasteurianum 28
pMS2pSK(+)::5,500-bp HindIII-BstEII fragment containing the dhaBCEF genes of C. freundii 41
pCS111pKK223-3::2,212-bp EcoI-HindIII fragment containing the His6-tagged dhaFG genes of C. freundii under control of the tac promoter 40
pCS120pSK(+)::2,731-bp EcoRI-HindIII fragment containing the His6-tagged dhaBCE genes of C. freundii under control of the lac promoter 40
pAK1pSTV28::8,536-bp BpiI-Bsp119I fragment containing the dhaDGHIKRT genes of C. freundiiThis study
pAK2pSK(+)::7,115-bp Bsp143I fragment containing dhaBCEFIT∗ genes derived from enrichment culturesThis study
pAK3pSK(+)::2,157-bp Bsp143I fragment containing dhaB∗CEF∗ genes derived from enrichment culturesThis study
pAK4pSK(+)::1,750-bp Bsp143I fragment containing pduC∗DE∗ genes derived from enrichment culturesThis study
pAK5pSK(+)::4,236-bp Bsp143I fragment containing the pduC∗DEGHJ genes derived from enrichment culturesThis study
pAK6pSK(+)::4,332-bp Bsp143I fragment containing parts of the C. freundii dha regulon (not further characterized)This study
pAK7pSK(+)::6,295-bp Bsp143I fragment the containing dhaBCEF∗GH∗IT genes derived from enrichment culturesThis study
pAK8pSK(+)::6,805-bp Bsp143I fragment containing parts of the C. freundii dha regulon (not further characterized)This study
pAK2.1pET100/D-TOPO::2,699-bp PCR product containing dhaBCE genes from pAK2 under control of the T7 promoterThis study
pAK3.1pET100/D-TOPO::2,699-bp PCR product containing a hybrid dhaB gene derived from C. freundii and pAK3 and the dhaCE genes of pAK3 under control of the T7 promoterThis study
pAK5.1pET101/D-TOPO::2,884-bp PCR product containing a hybrid pduC gene derived from S. enterica serovar Typhimurium and pAK5 and the pduDE genes of pAK5 under control of the T7 promoterThis study
pAK7.1pET101/D-TOPO::2,696-bp PCR product containing the dhaBCE genes of pAK7 under control of the T7 promoterThis study
  • a *, partial gene.