산업용 효모 Hybrid의 선별을 위한 우성선별표지로서의 Aureobasidin A 내성유전자의 이용
The Use of Aureobasidin A Resistant Gene as the Dominant Selectable Marker for the Selection of Industrial Yeast Hybrid
1
조회
0
다운로드
3
인용
0
참고문헌
키워드
초록
교배와 원형질체 융합을 통한 배수체인 야생형 산업균주의 개발을 위하여, hybrid의 선별표지로서 우성의 선별표지인 aureobasidin A 내성이 사용될 수 있는 지를 알아보고자하였다. 선별배지에서 aureobasidin A의 최적농도는 야생형균주인 경우 SD와 YPD 배지에서는 0.5 μg/mL 이상이었고,SG와 YPG에서는 0.2-0.3 μg/mL 이었다. 한편 호흡결여돌연변이주는 야생형 균주보다 훨씬 높은 농도의 aureobasidin A에도 내성이 있음을 나타내었다. 우리는 K114/YIP균주의전분분해 능력이 배수체 야생형 산업 균주에 전달 될 수 있는지를 이 방법을 통하여 관찰하였다. 반수체 영양요구성 균주 K114/YIP에 aureobasidin에 대한 내성을 부여하는pAUR112가 도입된 균주와 야생형 균주 KL 혹은 C6와의rare-mating 후 aureobasidin A 함유 배지에서 성장한 hybrid 를 분리할 수 있었다. Hybrid는 전분분해 능력을 함유하고있었을 뿐 아니라 두 양친의 특성을 동시에 지녔으며, 전자현미경 관찰 결과에서도 hybrid는 양친주의 특성을 모두 갖는 것으로 나타났다.
Abstract
For the strain improvement of the industrial polyploid yeast strain through hybridization and protoplast fusion, a dominant selection marker other than a recessive marker such as the auxotrophic marker was required for the selection of the resulting hybrids. In the present investigation, the aureobasidin A resistant gene was tested in relation to whether it can be used as the dominant selectable marker for the isolation of hybrids of the yeast Saccharomyces. The plasmid pAUR112, carrying the gene responsible for resistance to aureobasidin A, was introduced into the haploid yeast strain K114/YIp. From the rare-mating between polyploid C6 and haploid K114/YIp carrying pAUR112, many hybrids were obtained from the agar medium containing 0.5 μg/ml of aureobasidin A. The hybrids exhibited characteristics derived from both of the parental strains; and the cell sizes of the hybrids were larger than those of the parental strains. These results showed that the aureobasidin A resistant gene could be successfully used as the dominant selectable marker for the isolation of yeast hybrids resulting from rare-mating.
참고문헌 (20)
- Aeed, P. A., "Inhibition of inositol phosphorylceramidesynthase by the cyclic peptide aureobasidin", AntimicrobAgents Chemother, 53, pp.496~504 (2009)
- Cerantola, V., "Aureobasidin Aarrests growth of yeast cells through both ceramide intoxicationand deprivation of essential inisitolphosphorylceramides", Molecular Microbiology, 71, pp.1523~1537 (2009)
- Cerbón, J., "Inositol phosphoceramide synthase is a regulatorf intracellular levels of diacylglycerol and ceramide duringthe G1 to S transition in Saccharomyces cerevisiae", BiochemJ. 15, 388, pp.169~176 (2005)
- Cook, J. R., "Yeastartificial chromosome fragmentation vectors that utilizeURA3 selection", Genet. Anal. Tech. Appl, 10, pp.109~112 (1993)
- Cowart, L. A, "Yeast sphingolipids:recent developments in understanding biosynthesis, regulation,and function", Biochim Biophys Acta, 1771, pp.421~431 (2007)
- Dujon, B., "Complete DNA sequence of yeast chromosome XI", Nature, 369, pp.371~378 (1994)
- Endo, M., "Fungicidal action of aureobasidin A, a cyclic depsipeptideantifungal antibiotic, against Saccharomyces cerevisiae", Antimicrob. Agents. Chemother, 41, pp.672~676 (1997)
- Hashida-Okado, T., "AUR1, a novel gene conferring aureobasidinresistance on Saccharomyces cerevisiae : a study ofdefective morphologies in Aur1p-depleted cells", Mol. Gen.Genet, 251, pp.236~244 (1996)
- Heidler, S. A, "The AUR1 gene inSaccharomyces cerevisiae encodes dominant resistance toantifungal agent aureobasidin A (LY295337)", Antimicrob.Agents Chemother, 39, pp.2765~2769 (1995)
- Ikai, K., "Structure ofaureobasidin A.J.Antibiot.44:925-933", Structure ofaureobasidin A. J. Antibiot, 44, pp.925~933 (1991)
- Ito, M., "Transformation ofintact yeast cells treated with alkali cations or thiolcompounds", Agric. Biol. Chem, 48, pp.341~347 (1984)
- Jesch, S. A., "Interruption of inositol sphingolipidsynthesis triggers Stt4p-dependent protein kinase C signaling", J. Biol. Chem, 285, pp.41947~41960 (2010)
- Kim, T, "The construction of a stablestarch-fermenting yeast strain using genetic engineering andrare-mating", Appli. Biochemistry and Biotechnology, 59, pp.39~51 (1996)
- Kreger-van Rij, N. J. W, "The yeasts a taxonomicstudy" (1984), Elsevier Scinceublishers
- Rose, M., "Structure andfunction of the yeast URA3 gene : expression in Escherichiacoli", Gene, 29, pp.113~124 (1984)
- Saigal,D, "Review article : Yeast strain developmentfor ethanol production", Ind. J. Microbiol, 33, pp.159~168 (1993)
- Sherman, F., "Methods inYeast Genetics, Laboratory Course Manual, Cold SpringHarbor Laboratory" (1986), Cold Spring Harbor
- Spencer, J. F. T., "The use of mitochondrial mutant in hybridization ofindustrial yeast strains", Curr. Genet, 9, pp.649~652 (1985)
- Spencer, J. F. T, "Genetic improvementof industrial yeasts", Ann. Rev. Microbiol, 37, pp.121~142 (1983)
- Takesako, K., "Biological properties of aureobasidin A, a cyclic depsipeptideantifungal antibiotic", J. Antibiot, 46, pp.1414~1427 (1993)