Development of Primers for PCR Classification of Lactic Acid Bacteria in Kimchi Based on the Genes for Carbohydrate Metabolic Pathways

서지정보

ㆍ발행기관 : 한국유산균학회 ㆍ수록지정보 : Current Topic in Lactic Acid Bacteria and Probiotics / 2권 / 2호
ㆍ저자명 : Myung-Ki Lee, Hyo Jin Kim, Kyung Mo Song, Sung Hoon Yi, Young-Kyoung Rhee, Sung-Won Moon, Yu-Ri Kim

목차

Abstract
Introduction
Materials and Methods
Reference strains
DNA extraction from lactic acid bacteria
Development of identification primer
PCR amplification
Electrophoresis analysis of PCR product
Results and Discussion
Primer design based on metabolic enzyme gene sequences
PCR analysis of lactic acid bacteria with designedprimers
References

영어 초록

The aim of this study was to develop PCR primer sets for the rapid classification of lactic acid bacteria (LAB). Published gene sequences for LAB carbohydrate metabolic enzymes were collected from the NCBI GenBank, and 45 primer sets were designed using the gene sequences. Twelve species of LAB were used as reference strains: Lactobacillus brevis, Lactobacillus farciminis, Lactobacillus fermentum, Lactobacillus hilgardii, Lactobacillus plantarum, Weissella cibaria, Weissella confusa, Weissella kimchii, Leuconostoc citreum, Leuconostoc lactis, Pediococcus acidilactici, and Pediococcus pentosaceus. PCR amplification and gel electrophoresis were performed to verify the specificities of the designed primers. The results showed that the primer set No. 1 of 5'-aacaacaaaatcaccgcaca-3' & 5'- gtcgtcaatgttgtcgatgc-3' for phosphofructokinase amplified PCR products from 5 species of heterofermenter with different molecular weight depending on the genus. Primer set No. 8 of 5'-gcgtcgccgtctcg-3' & 5'- gcctgcggcttttcg-3' produced specific PCR products from three heterofermentative lactobacilli such as L. brevis, L. fermentum, and L. hilgardii. Primer set No. 18 of 5'-gtgacggtgctgtaggttca- 3' & 5'-gcagtcgcttacgccatatt-3' was specifically reacted to homofermentative species including L. farciminis, P. acidilactici, and P. pentosaceus. Hence, the primer sets which were developed in this study could be used as a tool for the classification and differentiation of homo- and hetero fermentative species in lactic acid fermented foods, like as kimchi.

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