TABLE 1.

Key parent plasmids and relevant derivatives of these plasmids used for the construction of A. vinelandii mutant strains

PlasmidaRelevant gene(s) clonedb,cVectorReference or source
pDB126418-kb BglII A. vinelandii chromosomal isc operon fragmentpUC119This study
    pDB12863.5-kb HindIII hscBA-fdx-iscX fragmentpUC119This study
    pDB12915.2-kb BspHI iscSUA-hscBA-fdx-iscX fragment with SbfI-ΔhscBA (aa HscB 78-HscA 442)pAra13This study
    pDB13165.2-kb BspHI iscSUA-hscBA-fdx-iscX fragment under PscrX controlpUC7This study
    pDB16083.2-kb PciI-BglII hscBA-fdx-iscX fragment under PscrX controlpUC7This study
pDB1307PCR-derived 3.82-kb scrX gene and promoter region (PscrX)pUC7This study
    pDB1309HincII-Knr from pUC-KAPA in EcoRV site of scrX genepUC7This study
    pDB1310MCS 1d replacing ΔPciI-KpnI fragment of scrX; introduces the following unique sites; PciI, XbaI, XhoI, and KpnIpUC7This study
    pDB13165.2-kb BspHI iscSUA-hscBA-fdx-iscX fragment under PscrX controlpUC7This study
    pDB1332MCS 2e replacing ΔPciI-KpnI in scrX; introduces the unique sites PciI, NruI, EcoRV, XhoI, BglII, XbaI, SphI, and KpnIpUC7This study
    pDB13356.1-kb SmaI-lacZ-kan fragment under PscrX controlpUC7This study
    pDB16083.2-kb PciI-BglII hscBA-fdx-iscX fragment under PscrX controlpUC7This study
    pDB16340.4-kb PciI-BamHI iscA fragment under PscrX controlpUC7This study
pDB9451-kb EcoRI fdx fragmentpUC11967
    pDB1016HincII-kan from pUC-KAPA in EcoRV site of fdx genepUC119This study
    pDB1550XhoI-Δfdx (Fdx aa 29-64)pUC119This study
pDB9332.5 kb EcoRI-SstI iscSUA fragmentpUC11967
    pDB954SalI-ΔiscS (IscS aa 14-182)pUC119This study
    pDB983NdeI-BamHI iscSUA fragmentpT7-7This study
    pDB1058Site-directed substitution; iscU codon 39, GCC (IscUD39A)pUC119This study
    pDB1209Site-directed substitution; iscS codon 325, GCC (IscSC325A)pUC119This study
    pDB1215Site-directed substitution; iscA codon 99, GCC (IscAC99A)pUC119This study
    pDB1216Site-directed substitution; iscA codon 101, GCC (IscAC101A)pUC119This study
    pDB1227Site-directed substitution; iscU codon 106, GCG (IscUC106A)pUC119This study
    pDB1228Site-directed substitution; iscU codon 63, GCC (IscUC63A)pUC119This study
    pDB1236Site-directed substitution; iscA codon 36, GCG (IscAC36A)pUC119This study
    pDB1350ΔiscU (IscU aa 31-117) from Bal31 exonuclease digestion at DraIIIpT7-7This study
    pDB1351ΔiscUA (IscU aa 34-IscA 9) from Bal31 exonuclease digestion at SexAI in iscApT7-7This study
    pDB1391ΔiscA (IscA aa 8-125) from Bal31 exonuclease digestion at SexAIpT7-7This study
    pDB1404Site-directed substitution; iscU codon 37, GCG (IscUC37A)pUC119This study
    pDB1538Site-directed substitution; iscU codon 105, GCC (IscUH105A)pUC119This study
    pDB1546Site-directed substitution; iscU codon 103, GCG (IscUK103A)pUC119This study
Other
    pDB528recA::Tn10 (Knr)pSUP10263
    pDB13752.6-kb PstI-gn from pWKR202I in PstI site of PCR-derived recA genepUC7This study
    pDB14002.6-kb blunt-ended BamHI-gn from pWKR2021 in StuI site of PCR-derived scrR genepUC19This study
    pDB14686.1-kb SmaI-lacZ-kan from pLKC482 in AleI site of PCR-derived hscA genepT7-7This study
  • a Key parent plasmids are written in boldface type. Plasmid derivatives are listed directly below the parent.

  • b Sizes of gene deletions are described as amino acid (aa) deletions.

  • c Descriptions of site-directed amino acid substitutions include the corresponding DNA codon change.

  • d A 45-bp multiple cloning site (see Materials and Methods for sequence).

  • e A 35-bp multiple cloning site (see Materials and Methods for sequence).