Differentiation between brewing and non-brewing yeasts using a combination of PCR and RFLP.
In order to differentiate brewing from non-brewing yeasts, a specific polymerase chain reaction (PCR) which targeted the open reading frame of FLO1 was employed. Non-brewing yeasts include 'non-brewing Saccharomyces yeasts' and 'non-Saccharomyces yeasts'. The molecular sizes of the PCR products differed between brewing and non-brewing Saccharomyces yeasts. No FLO1 PCR products were obtained from non-Saccharomyces yeasts. Specific PCR, using oligonucleotide primers that targeted the region between the 5S and 26S rRNA genes, could be used to differentiate brewing yeasts from some non-brewing yeasts. These PCR products were digested with restriction enzymes, Scr FI and Msp I. Different restriction profiles were obtained from brewing and non-brewing yeasts which could not be differentiated using specific PCR of rDNA. These results suggest that it is possible to identify brewing from non-brewing yeasts using specific PCR of FLO1 and rDNA, and detection of restriction fragment polymorphism of rDNA.
Yamagishi
H
H
Brewing Research & Development Laboratory, Asahi Breweries Ltd, Ibaraki, Japan. hiromi.yamagishi@asahibeer.co.jp
Otsuta
Y
Y
Funahashi
W
W
Ogata
T
T
Sakai
K
K
D001515:Beer / Q000382:microbiology*; D015139:Blotting, Southern; D017931:DNA Primers; D004271:DNA, Fungal / Q000032:analysis; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000302:isolation & purification*; D013045:Species Specificity
D017931:DNA Primers; D004271:DNA, Fungal