Authors: Valcheva, R.; Kabadjova, P.; Rachman, C.¹; Ivanova, I.²; Onno, B.¹; Prévost, H.¹; Dousset, X.¹

Source: Journal of Applied Microbiology, Volume 102, Number 1, January 2007 , pp. 290-302(13)

Publisher: Wiley-Blackwell

Abstract:

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-CeuI 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.

Keywords: 16S-23S rDNA intergenic spacer regions; Lactobacillus sanfranciscensis; rrn operons; species-specific PCR; tRNA

Document Type: Research article

DOI: http://dx.doi.org/10.1111/j.1365-2672.2006.03039.x

Affiliations: 1:  Laboratoire de Microbiologie Alimentaire et Industrielle, QM2A, ENITIAA, Nantes Cedex 3, France 2:  Department of General and Industrial Microbiology, Faculty of Biology, University of Sofia, Sofia, Bulgaria

Publication date: 2007-01-01

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