乳酸菌における多剤耐性遺伝子(lmrA・lmrP)の検索とクローニング

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  • Detection and Cloning of Multidmg Resistance Genes (lmrA・lmrP) in Several Lactic Acid Bacteria
  • ニュウサンキン ニ オケル タザイ タイセイ イデンシ lmrA lmrP ノ ケンサク ト クローニング

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The genes, lmrA and lmrP, are identified as multidrug resistant genes in Lactococcus lactis subsp lactis MG1363, and these genes express LmrA and LmrP which are multidrug transporters (Konings et al., 1994-2000^<1-14)>). Multidrug transporters are a representative mechanism of multidrug resistance in bacteria and mammals. The amino acid sequence of LmrA showed 30% homology to the N- and C-terminal halves of the human multidrug resistant P-glycoprotein MDR1. And especially the sequence identities of ATP binding cassette domain of these protein are high (48% and 43% identical, respectively). In this study, attempts were made to confirm the presence of multidrug resistant genes in Lactococcus and Lactobacillus, which are lactic acid bacteria. The complete genomic DNA sequences of both lmrA and lmrP were isolated from 10 strains of Lactococcus lactis subsp. lactis by PCR amplification, whereas the presence of the homologous genomic DNA sequences genes was not confirmed in Lactococcus lactis subsp. cremoris and Lactobacillus. Vector pCAGGS (Niwa et al., 1991^<15)>) was used as the expression vector for lmrA in mammalian cells in this study. The PCR products of lmrA was cloned into pCAGGS in XhoI site, and pCAGGS-lmrA (7,873 bp) was constructed.

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