Random amplified polymorphic DNA-PCR based cloning of markers to identify the beer-spoilage strains of Lactobacillus brevis, Pediococcus damnosus, Lactobacillus collinoides and Lactobacillus coryniformis.
OBJECTIVE
Beer-spoilage ability of lactic acid bacteria such as Lactobacillus brevis is a strain-dependent phenomenon in which the mechanism has not yet been completely clarified. In order to systematically identify genes that contribute to beer-spoilage, large-scale random amplified polymorphic DNA (RAPD)-based cloning methods was carried out.


RESULTS
A systematic RAPD polymerase chain reaction (PCR) analysis using 600 primers was performed on beer-spoilage and on nonspoilage strains of L. brevis. Among 600 primers, three were found to amplify a single locus highly specific to beer-spoilage strains. DNA sequencing of this locus revealed a three-part operon encoding a putative glycosyl transferase, membrane protein and teichoic acid glycosylation protein. PCR analysis of typical beer-spoilage lactic acid bacteria suggested that this locus is highly specific to beer-spoilage strains.


CONCLUSIONS
The cloned markers are highly specific to identify the beer-spoilage strains not only in L. brevis but also in Pediococcus damnosus, Lactobacillus collinoides and Lactobacillus coryniformis.


CONCLUSIONS
This paper proves that RAPD-PCR is an efficient method for cloning the strain-specific genes from bacteria. The markers described here is one of the most useful tools to identify the beer-spoilage strains of lactic acid bacteria.
Fujii
T
T
Central Laboratories for Key Technology, Kirin Brewery Co., Ltd, Fukuura, Kanazawa, Yokohama, Kanagawa, Japan. tfuiji@kirin.co.jp
Nakashima
K
K
Hayashi
N
N
D001515:Beer; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000032:analysis*; D005506:Food Contamination; D005516:Food Microbiology; D007778:Lactobacillus / Q000235:genetics*; D010376:Pediococcus / Q000235:genetics*; D019105:Random Amplified Polymorphic DNA Technique
D004269:DNA, Bacterial