Analysis of Bacterial Communities in Kusaya Gravy by Denaturing Gradient Gel Electrophoresis of PCR-Amplified Ribosomal DNA Fragments.
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概要
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We analyzed the microbial communities in <I>kusaya</I> gravy obtained from three manufacturers by a combination of cultivating and molecular typing methods, PCR-RFLP and PCRDGGE. Viable counts were 1.2×10<SUP>8</SUP>-1.5×10<SUP>9</SUP>/g on TSA medium, BP<SUB>5</SUB>G medium and ABCM medium. With PCR-RFLP, forty isolates obtained from BP<SUB>5</SUB>G medium and ABCM medium were classified into six to nine groups. Representative strains of the isolates were identified as <I>Pseudomonas, Marinobacter, Peptostreptococcus, Enterococcus</I> etc. with 16S rDNA sequencing. DGGE analysis of 16S rDNA fragments amplified directly from each of three kinds of <I>kusaya</I> gravy showed a similar banding pattern each other. Several main fragments were recovered from DGGE-gel and were cloned and sequenced. The existence of <I>Bacteroides, Flavobacterium, Fusobacterium, Eggerthela lenta</I> and <I>Clostridium</I> was indicated from the analysis of fragments. Therefore, there are great differences between the dominant organisms obtained by culturing and by PCR-DGGE, suggesting that unculturable organisms prevail in <I>kusaya</I> gravy.
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