Open AccessMolecular characterization of semi-hard homemade cheese microflora
Author(s) -
Branko Jovčić,
Nataša Golić,
Milan O. Kojić,
Ljubiša Topisirović
Publication year - 2005
Publication title -
acta veterinaria
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.308
H-Index - 17
eISSN - 1820-7448
pISSN - 0567-8315
DOI - 10.2298/avb0506511j
Subject(s) - biology , 16s ribosomal rna , bacteria , rna , dna , oligonucleotide , gene , dna extraction , polymerase chain reaction , microbiology and biotechnology , food science , genetics
This decade has shown an impressive development in the application of molecular techniques based on 16S and 23S rRNA genes to study the microbial diversity in various ecosystems. Microflora of semi-hard homemade cheese was examined in this work. We developed a novel technique for DNA extraction (a bead beating based method) due to high fat content of this cheese. Rapid extraction of DNA from cheese microflora enabled a molecular identification of the LAB (Lactic Acid Bacteria) strains based on PCR amplification of 16S RNA coding sequences. The specific primers for 76S RNA gene of lactobacilli were used for amplification. The PCR reaction was performed at lower temperature, where the specificity of the annealing reaction was reduced and lactococcal sequences of 16S RNA genes were also amplified. The results of RFLP analysis revealed that the microflora of Doboj homemade cheese encompases mostly lactococci