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A rapid PCR procedure for the specific identification of Lactobacillus sanfranciscensis , based on the 16S‐23S intergenic spacer regions
Author(s) -
Valcheva R.,
Kabadjova P.,
Rachman C.,
Ivanova I.,
Onno B.,
Prévost H.,
Dousset X.
Publication year - 2007
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 - 10.1111/j.1365-2672.2006.03039.x
Subject(s) - biology , spacer dna , 23s ribosomal rna , operon , internal transcribed spacer , ribosomal rna , genetics , ribosomal intergenic spacer analysis , microbiology and biotechnology , gene , rna , ribosome , escherichia coli
Aims: The organization of ribosomal RNA ( rrn ) operons in Lactobacillus sanfranciscensis was studied in order to establish an easy‐to‐perform method for identification of L. sanfranciscensis strains, based on the length and sequence polymorphism of the 16S‐23S rDNA intergenic spacer region (ISR). Methods and Results: PCR amplification of the 16S‐23S rDNA ISRs of L. sanfranciscensis gave three products distinguishing this micro‐organism from the remaining Lactobacillus species. Sequence analysis revealed that two of the rrn operons were organized as in previously reported lactobacilli: large spacer (L‐ISR), containing tRNA Ile and tRNA Ala genes; small spacer (S‐ISR) without tRNA genes. The third described spacer (medium, M‐ISR), original for L. sanfranciscensis , harboured a tRNA‐like structure. An oligonucleotide sequence targeting the variable region between tDNA Ile and tDNA Ala of L. sanfranciscensis L‐ISR was approved to be suitable in species‐specific identification procedure. Analysis by pulse‐field gel electrophoresis of the chromosomal digest with the enzyme I‐ Ceu I showed the presence of seven rrn clusters. Lactobacillus sanfranciscensis genome size was estimated at c. 1·3 Mb. Conclusions: Direct amplification of 16S‐23S ISRs or PCR with specific primer derived from L‐ISR showed to be useful for specific typing of L. sanfranciscensis . This was due to the specific rrn operon organization of L. sanfranciscensis strains. Significance and Impact of the Study: In this paper, we have reported a rapid procedure for L. sanfranciscensis identification based on specific structures found in its rrn operon.