Cloning of a novel specific SCAR marker for species identification in Lactobacillus pentosus

Mol Cell Probes. 2014 Aug;28(4):192-4. doi: 10.1016/j.mcp.2014.03.003. Epub 2014 Mar 24.

Abstract

Identifying Lactobacillus species using only phenotypic and genotypic (16S rDNA sequence analysis) techniques yields inaccurate results. The objective of this study was to develop species-specific primers based on randomly amplified polymorphic DNA (RAPD) fingerprinting to distinguish species within the closely related Lactobacillus plantarum group. One of these primers, OPD-3, produced a species-specific band that was found only in the tested Lactobacillus pentosus. This specific fragment was isolated from agarose gel and ligated into a vector for DNA sequencing. A pair of primers, SpOPD3Lpen-F1/R1, that were highly specific sequence-characterized-amplified-regions (SCARs) were designed according to the nucleotide sequences of the specific RAPD marker. These primers were used for PCR analysis of the template DNA of the Lactobacillus strains, and a single 542 bp species-specific band was found only in L. pentosus. Using PCR, a novel species-specific primer pair is shown to rapidly, accurately and effectively distinguish L. pentosus from other species in the L. plantarum group of probiotic bacteria.

Keywords: Lactobacillus pentosus; RAPD fingerprinting; Species identification; Species-specific PCR.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Cloning, Molecular
  • DNA Primers / genetics*
  • DNA, Bacterial / genetics
  • Genetic Markers
  • Lactobacillus / classification*
  • Lactobacillus / genetics*
  • Lactobacillus / isolation & purification
  • Molecular Sequence Data
  • Random Amplified Polymorphic DNA Technique
  • Species Specificity

Substances

  • DNA Primers
  • DNA, Bacterial
  • Genetic Markers