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Characterization of Lactobacillus sake isolates from dry‐cured sausages by restriction fragment length polymorphism analysis of the 16S rRNA gene
Author(s) -
Sanz Y.,
Hernández M.,
Ferrús M.A.,
Hernández J.
Publication year - 1998
Publication title -
journal of applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 1364-5072
DOI - 10.1046/j.1365-2672.1998.00387.x
Subject(s) - hindiii , ecori , biology , restriction fragment length polymorphism , restriction enzyme , genetics , ribotyping , 16s ribosomal rna , microbiology and biotechnology , southern blot , genomic dna , restriction fragment , complementary dna , terminal restriction fragment length polymorphism , dna , genotype , gene
Lactobacillus sake strains originally isolated from dry‐fermented sausages were characterized by phenotypic and genotypic methods, including DNA‐DNA hybridization, restriction fragment length polymorphism (RFLP), and 16S rDNA sequencing analysis, in order to establish their taxonomic position and relation to well defined reference species. Initially, isolates of Lact. sake showing a characteristic phenotype (melibiose‐positive, maltose‐ and arabinose‐negative) were identified by DNA‐DNA hybridization. Subsequently, RFLP studies using Eco RI and Hin dIII as restriction enzymes, and cDNA from Escherichia coli or 16S rDNA from Lact. sake strains as probes, showed distinct polymorphism levels. Thus, Eco RI‐digested DNA probed with cDNA from E. coli disclosed the presence of a unique cluster for the meat isolates tested, allowing their differentiation from the reference type strain. When Hin dIII‐digested DNA was hybridized with the cDNA probe, strain‐specific patterns were obtained, showing a higher discrimination power. Considerable strain differentiation was also observed when Eco RI and Hin dIII digests were hybridized with 16S rDNA probes. Finally, sequence analysis of the 16S rDNA from one isolate also revealed a certain degree of genetic variability with respect to the reference strain of Lact. sake .