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Cascade cell lyses and DNA extraction for identification of genes and microorganisms in kefir grains

Kowalczyk, Magdalena and Kołakowski, Piotr and Radziwill-Bienkowska, Joanna M. and Szmytkowska, Agnieszka and Bardowski, Jacek K. (2012) Cascade cell lyses and DNA extraction for identification of genes and microorganisms in kefir grains. Journal of Dairy Research, 79 (01). pp. 26-32.

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Official URL: http://journals.cambridge.org/action/displayJourna...

Abstract

Kefir is a dairy product popular in many countries in Central Europe, especially in Poland and other countries of Eastern and Northern Europe. This type of fermented milk is produced by a complex population of symbiotic bacteria and yeasts. In this work, conditions for DNA extraction, involving disruption of kefir grains and a cascade of cell lysis treatments, were established. Extraction procedure of total microbial DNA was carried out directly from fresh kefir grains. Using different lysis stringency conditions, five DNA pools were obtained. Genetic diversity of DNA pools were validated by RAPD analysis, which showed differences in patterns of amplified DNA fragments, indicating diverse microbial composition of all the analysed samples. These DNA pools were used for construction of genomic DNA libraries for sequencing. As much as 50% of the analysed nucleotide sequences showed homology to sequences from bacteria belonging to the Lactobacillus genus. Several sequences were similar to sequences from bacteria representing Lactococcus, Oenococcus, Pediococcus, Streptococcus and Leuconostoc species. Among homologues of yeast proteins were those fromCandida albicans, Candida glabrata, Kluyveromyces lactis and Saccharomyces cerevisiae. In addition, several sequences were found to be homologous to sequences from bacteriophages.

Item Type:Article
Additional Information:This paper has been accepted for publication and will appear in a revised form, subsequent to peer review and/or editorial input by Cambridge University Press, in Journal of Dairy Research published by Cambridge University Press, together with a copyright notice in the name of the copyright holder (Cambridge University Press)
Subjects:Q Science > QR Microbiology
Divisions:Department of Microbial Biochemistry
ID Code:215
Deposited By: Dr Magdalena Kowalczyk
Deposited On:15 Mar 2012 08:22
Last Modified:08 Mar 2018 15:33

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