title: Intraspecies discrimination of Lactobacillus paraplantarum by PCR.
abstract: Lactobacillus paraplantarum is a species phenotypically close to Lactobacillus plantarum. Several PCR methods were evaluated to discriminate L. paraplantarum strains and among them, a PCR using an enterobacterial repetitive intergenic consensus (ERIC) sequence differentiated L. paraplantarum from other Lactobacillus species. In addition, a combination of ERIC and random amplified polymorphic DNA (RAPD) analysis distinguished among seven strains of L. paraplantarum tested. ERIC-PCR profiles showed several strain-specific DNA fragments in L. paraplantarum, among them, a 2.2-kb ERIC marker, termed LpF1, found to be specific to strain FBA1, which improved the skin integrity in an animal model. The LpF1 encodes three proteins similar to Lactobacillus fermentum AroA, TyrA, and AroK, which are involved in the shikimate pathway. A primer pair specific to FBA1 based on the internal sequence of LpF1 amplified a 950-bp FBA1-specific fragment LpF2. Southern blot analysis of Dra I-digested genomic DNA of L. paraplantarum strains using LpF2 as a probe showed that LpF2 is distinctive of strain FBA1 among 16 L. paraplantarum strains. Because both ERIC- and RAPD-PCR are fast and technically simple methods, they are useful for the rapid discrimination of L. paraplantarum strains and for the development of new strain-specific DNA markers for identifying industrially important strains.
authors.affiliation: National Institute of Livestock and Grassland Science, Tsukuba, Ibaraki, Japan.
authors.forename: Shinichi
authors.initials: S
authors.lastname: Saito
authors.forename: Miho
authors.initials: M
authors.lastname: Kobayashi
authors.forename: Hiromi
authors.initials: H
authors.lastname: Kimoto-Nira
authors.forename: Reiji
authors.initials: R
authors.lastname: Aoki
authors.forename: Koko
authors.initials: K
authors.lastname: Mizumachi
authors.forename: Satoshi
authors.initials: S
authors.lastname: Miyata
authors.forename: Katsue
authors.initials: K
authors.lastname: Yamamoto
authors.forename: Yasushi
authors.initials: Y
authors.lastname: Kitagawa
authors.forename: Chise
authors.initials: C
authors.lastname: Suzuki
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics; D015373:Bacterial Typing Techniques / Q000379:methods*; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D007778:Lactobacillus / Q000145:classification* / Q000235:genetics*; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA
chemical_list: D001426:Bacterial Proteins; D017931:DNA Primers; D004269:DNA, Bacterial; D015345:Oligonucleotide Probes