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KMID : 0903520060490030186
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Journal of the Korean Society of Agricultural Chemistry and Biotechnology
2006 Volume.49 No. 3 p.186 ~ p.191
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Electrofusion of Yeast Cells and Their Genetic Analysis Using RAPD-PCR
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Kim Seung
Kim Jae-Sung
Sapkota Kumar
Park In-Sung
Cho Moon-Gu
Park Yeal
Chun Hong-Sung
Choi Bong-Suk
Park Se-Eun
Choi Han-Seok
Kim Myung-Kon
KIm Sung-Jun
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Abstract
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In the present study, an attempt has been made to produce hybrid yeast strains of different useful and dominant characteristics. The hybrid yeast strains were produced by electrofusion and their genetic analysis were performed by RAPD-PCR (random amplified polymorphic DNA-polymerase chain reaction). The protoplast of Saccharomyces cerevisiae KCTC 7904 and Zygosaccharomyces rouxii KCTC 7966 were obtained above 92% when treated with lyticase at 30oC for 60~90 min after the pretreatment of 1~2% 2-mercaptoethanol at 30oC for 15~20 min. The fusant was produced from paired protoplast stage under the electric pulse at high frequency conditions (1.5 MHz/ 50 pV, 615 V/256 μsec) within glass-platinum made electrofusion chamber. Changes in RAPD patterns in mother cells and hybrid cells proved that the fusant contains two types of yeast gene originated from its parent. Furthermore, fermentation characters exhibits by the fusant cell confirmed its genetic changes. These results suggest that genetically stable hybrid yeast strains of economic importance can be produced by electrofusion technique and these electrofused yeast cells have an enormous impact in biotechnology and biomedicine.
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KEYWORD
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Yeast, Protoplast, Electrofusion, Fusant, Random amplified polymorphic DNA-polymerase chain reaction
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