TABLE 1.

L. biflexa mutants isolated by random insertional mutagenesis

Clone (mutant)Himar1 insertion site (TA)aBLAST results (% identity)bPutative interrupted ORFcAuxotrophic phenotype
Casamino Acids screen
    BtgacgtcttTAtttgatattL. interrogans indole-3-glycerol phosphate synthase (54)trpC (Prom) AY828565Tryptophan
    CtaggttatgaTAttgaggaattL. interrogans signal transduction histidine kinase (43)hkld (5′) AY828564Glutamate
    DaacaaagagTAtgcgggcgtBradyrhizobium japonicum glutamate synthase (53)gltB (5′) AY828568Glutamate
    EcaaggtatgTAtgatcctgcB. japonicum glutamate synthase (53)gltB (5′) AY828568Glutamate
    GcatagtacaTAtgaggtgaaL. interrogans anthranilate synthase component I (85)trpE (3′) AY828566Tryptophan
Hemin screen
    LtgatgaacgTAggtctgcaaL. interrogans iron(III) dicitrate TonB-dependent receptor (49)fecA (3′) AY744288Hemin
    PgacagacaaTAtggaaatctL. interrogans iron(III) dicitrate TonB-dependent receptor (49)fecA (5′) AY744288Hemin
    QtggaatgttTAataacaatcL. interrogans iron(III) dicitrate TonB-dependent receptor (49)fecA (5′) AY744288Hemin
    VagtaaaaaaTAaacttttgaL. interrogans ferrous iron transport protein B (52)feoB (3′) AY744289Hemin
    WctttgcattTAcaagaacaaL. interrogans ferrous iron transport protein B (52)feoB (5′) AY744289Hemin
  • a Himar1 flanking sequences (lowercase) were deduced by ligation-mediated PCR as described in Materials and Methods. Each clone contains a unique inserted transposon, as demonstrated by Southern analysis.

  • b Best BLAST score of the putative ORF interrupted by the Himar1 insertion.

  • c In parentheses, location of Himar1 in the coding sequence; Prom, location of Himar1 in the trpC promoter region. The GenBank accession number of the L. biflexa locus is indicated.

  • d hkl, histidine kinase from L. biflexa.