Isolation of a haploid from an industrial Chinese rice wine yeast for metabolic engineering manipulation

摘要: The complex metabolic processes of yeast influence wine fermentation and therefore the quality wine. Wine yeasts, owing to their being typically prototrophic often polyploid, have been restricted in terms exploiting classical recombinant genetic techniques improve characteristics. To overcome this problem, haploids isolated from a commercial Chinese rice strain N85, by disruption HO gene. In study, Cre–loxP system removable G418r marker were used construct an cassette. Most heterologous sequences constructed cassette successfully excised genome loop-out KanMX gene, through induced expression Cre recombinase. removal resistant ensures biological safety strains. As expected, no difference capacity between parental haploid strains was seen. present work reports construction touchdown polymerase chain reaction its application with for isolation broaden physiological investigations industrial applications. Copyright © 2013 Institute Brewing & Distilling