Physical and genetic map of the Lactobacillus sakei 23K chromosome

• Published in: Microbiology (Society for General Microbiology) Vol. 148; no. 2; pp. 421 - 431
• Main Authors: Dudez, Anne-Marie, Chaillou, Stephane, Hissler, Lionel, Stentz, Regis, Champomier-Verges, Marie-Christine, Alpert, Carl-Alfred, Zagorec, Monique
• Format: Journal Article
• Language: English
• Published: Reading Soc General Microbiol 01-02-2002; Society for General Microbiology; Microbiology Society
• Subjects: Bacterial Proteins - genetics; Bacteriology; Base Sequence; Biological and medical sciences; Chromosome Mapping; Chromosomes, Bacterial - genetics; DNA Probes - genetics; DNA, Bacterial - genetics; Electrophoresis, Gel, Pulsed-Field; Fundamental and applied biological sciences. Psychology; Genes, Bacterial; Genetic Markers; Genetics; Genome, Bacterial; Lactobacillus - genetics; Lactobacillus sakei; Life Sciences; Microbiology; Microbiology and Parasitology; Multigene Family; Nucleic Acid Hybridization; Physical Chromosome Mapping; Polymerase Chain Reaction; Restriction Mapping; rrn gene; rRNA 23S; Genetic mapping; Lactic acid bacteria; Pulsed field electrophoresis; Genetic marker; Ribosomal DNA; Bacteria; Lactobacillaceae; Physical map; meat; PFGE; lactic acid bacteria
• ISSN: 1350-0872; 1465-2080
• DOI: 10.1099/00221287-148-2-421

Unité Flore Lactique et Environnement Carné, INRA, Domaine de Vilvert, F-78350 Jouy-en-Josas, France 1
Author for correspondence: Monique Zagorec. Tel: [phone]. Fax: [phone]. e-mail: zagorec{at}diamant.jouy.inra.fr
The Lactobacillus sakei 23K chromosome was analysed by pulsed-field gel electrophoresis after digestion with the restriction enzymes Asc I, Not I and Sfi I. The chromosome size was estimated to be 1845±80 kb. The use of I- Ceu I, specific for rrn genes encoding 23S rRNAs, showed that seven rrn loci were present, on 40% of the chromosome. The seven rrn clusters were mapped and their orientation was determined, allowing the position of the replication origin to be estimated. Partial I- Ceu I digestions were used to construct a backbone and the different restriction fragments obtained with Asc I, Not I and Sfi I were assembled to a physical map by Southern hybridization. Eleven L. sakei gene clusters previously identified were mapped, as well as 25 new loci located randomly on the chromosome and 11 regions flanking the rrn gene clusters. A total of 47 clusters were thus mapped on L. sakei chromosome. The new loci were sequenced, allowing the identification of 73 complete or incomplete coding sequences. Among these 73 new genes of L. sakei , the function of 36 could be deduced from their similarity to known genes described in databases. However, 10 genes had no homologues, 10 encoded proteins similar to proteins of unknown function and 17 were similar to hypothetical proteins.
Keywords: lactic acid bacteria, meat, PFGE Abbreviations: LM-PCR, ligation-mediated polymerase chain reaction
The GenBank accession numbers for the sequences reported in this paper are given in Table 2 and the legend to Fig. 3.
a Present address: Institute of Food Research, Norwich Research Park, Norwich NR4 7UA, UK.
b Present address: Unité Ecologie et Physiologie du Système Digestif, INRA, Domaine de Vilvert, F78350 Jouy-en-Josas, France.