PCR-Based Differentiation and Homology of Brewing and Type Strains of the Genus Saccharomyces
Wojciech Barszczewski1,2 and Malgorzata Robak1
1 Department of Biotechnology and Food Microbiology, Faculty ofFood Science, Agricultural University of Wroclaw, Norwida 25,50-375 Wroclaw, Poland.
2 Corresponding author. E-mail: barszcz4@o2.pl
J. Inst. Brew. 112(2), 165–172, 2006 | VIEW ARTICLE
ABSTRACT
Restriction fragment length polymorphism (RFLP) patterns of PCR-amplified ribosomal RNA gene fragments (rDNA) and randomly amplified polymorphic DNA (RAPD) were applied for the analysis of 15 brewing and 6 related yeast strains of the ge-nus Saccharomyces. One five-base (ScrFI) and two four-base cutting (HaeIII, MspI) restriction enzymes were used. The prim-ers 21 and M13 core sequence were selected for RAPD analysis. PCR-RFLP rDNA analysis with HaeIII, ScrFI and MspI differentiated the strains tested into four, five and four types of pat-terns, respectively and the analyses of the profiles showed 100% homology, between the yeast strains. One strain was an exception. Homological groups were observed for strains used in breweries globally, from a local production strain and from the isolates identified as S. cerevisiae. Using RAPD analysis, and according to discrete differences in the profiles, it was possible to divide twenty one strains into 15 and 20 groups with primer 21 and M13 respectively. RFLP-PCR rDNA analysis was used to show similarities in closely related brewing strains, while RAPD analysis was used for differentiation of strains.
Key words:
brewing, PCR, RAPD, rDNA, RFLP, yeast.
Publication no. G-2006-0630-426 ©2006 The Institute & Guild of Brewing