E AKEGG pathway (or putative pathway)c Wax ester biosynthesis Wax

E AKEGG pathway (or putative pathway)c Wax ester biosynthesis Wax ester biosynthesis Wax ester biosynthesis Unknown Ribosome Homologous recombinationProtein/nucleotide accession no. YP_959769 YP_959486 YP_960668 YP_958650 NR_027551 YP_Provided as a uncomplicated reference for the gene products (or genes) shown in the figures. The complete name of the gene item, based on the NCBI reference number or preceding annotations. c Possible pathway in which the gene item is involved, from the KEGG or the pathway shown in Fig. 1. d Even though annotated here as fatty aldehyde reductase, we also note that this gene solution has been reported to possess fatty acyl-CoA reductase and fatty acyl-ACP reductase activities by other folks (4, 5). e 3 copies on the 16S rRNA gene are located in the genome.aem.asm.orgApplied and Environmental MicrobiologyFatty Alcohol Biosynthesis in MarinobacterTABLE two Important parent plasmids and their relevant derivatives made use of for the construction of Marinobacter aquaeolei VT8 manipulated strainsPlasmida pBBR1MCS-2 pSMV3 pBB053 pBB114 pBBTET3 pPCRKAN4 pPCRMOB4 pPCRSACB6 pPCRSACB7 pPCRWEK4 pPCRWEK5 pPCRWEK12 pPCRWEK14 pPCRWEK20 pPCRWEK26 pPCRWEK27 pPCRWEK29b,c pPCRWEK32 pPCRWEK33c pPCRWEK48 pPCRWEK50b,ca bRelevant gene cloned or plasmid manipulation(s) Plasmid containing mobilization element Plasmid containing sacB gene NdeI website removed from pUC19 by silent mutation Replaced pUC19 Amp resistance with Kan resistance cassette from pUC4K, then removed NsiI and HindIII web-sites from cassette by silent mutations Replaced pUC19 Amp resistance with Tet resistance cassette from pRK415 Cloned Kan cassette from pBBR1MCS-2 into pBBTET3 Moved mobilization element from pBBR1MCS-2 into pUC19 sacB gene from pSMV3 cloned into pBB053, and after that EcoRI, HindIII, XbaI, and KpnI sites had been removed by site-specific mutagenesis with silent mutations Moved sacB gene cassette from pPCRSACB6 to pBBTET3 Derivative of pBB053 for gene insertions Moved mobilization element from pPCRMOB4 into pPCRWEK4 Cloned acrB and flanking regions from M.Mirogabalin aquaeolei VT8 genome with primers BBP1477 and BBP1478 into pBBTET3 EcoRI and XbaI internet sites Performed PCR with primers to take away acrB from pPCRWEK12, leaving flanking regions and adding BamHI website Cloned farA and flanking regions from M. aquaeolei VT8 genome with primers BBP1522 and BBP1523 into pBBTET3 EcoRI and XbaI web pages Moved acrB flanking segments fragment from pPCRWEK14 into pPCRWEK5 Performed PCR with primers to remove farA from pPCRWEK20, leaving flanking regions and adding BamHI site Moved Kan cassette from pPCRKAN4 into BamHI-cut pPCRWEK26 Moved farA flanking segments fragment from pPCRWEK27 into pPCRWEK5 Moved Kan cassette from pPCRKAN4 into BamHI-cut pPCRWEK32 Moved sacB cassette from pPCRSACB7 into KpnI- and HindIII-cut pPCRWEK32 Moved Kan cassette cut with BamHI from pPCRKAN4 into BglII-cut pPCRWEKVectorReference and/or supply 14 9 This study pUC4K (22) pRK415 (23) This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This studypUC19 pUC19 pUC19 pBBTET3 pUC19 pBB053 pBBTET3 pBB053 pBB053 pBBTET3 pBBTET3 pBBTET3 pBB053 pBBTET3 pBB053 pBB053 pBB053 pBB053 pBBThe sequences of all plasmids utilized within this study are accessible upon request.Blonanserin Plasmid map can also be offered in Fig.PMID:23376608 2. c Plasmids shown in bold are completed vectors employed to transform M. aquaeolei VT8.a separate sample was taken for isolation of cells for quantification of.