Simultaneous and Sequential Integration by Cre/loxP Site-Specific Recombination in Saccharomyces cerevisiae
1
조회
0
다운로드
3
인용
0
참고문헌
키워드
초록
A Cre/loxP-δ-integration system was developed to allow sequential and simultaneous integration of a multiple gene expression cassette in Saccharomyces cerevisiae. To allow repeated integrations, the reusable Candida glabrata MARKER (CgMARKER) carrying loxP sequences was used, and the integrated CgMARKER was efficiently removed by inducing Cre recombinase. The XYLP and XYLB genes encoding endoxylanase and β-xylosidase, respectively, were used as model genes for xylan metabolism in this system, and the copy number of these genes was increased to 15.8 and 16.9 copies/cell, respectively, by repeated integration. This integration system is a promising approach for the easy construction of yeast strains with enhanced metabolic pathways through multicopy gene expression.
Abstract
A Cre/loxP-δ-integration system was developed to allow sequential and simultaneous integration of a multiple gene expression cassette in Saccharomyces cerevisiae. To allow repeated integrations, the reusable Candida glabrata MARKER (CgMARKER) carrying loxP sequences was used, and the integrated CgMARKER was efficiently removed by inducing Cre recombinase. The XYLP and XYLB genes encoding endoxylanase and β-xylosidase, respectively, were used as model genes for xylan metabolism in this system, and the copy number of these genes was increased to 15.8 and 16.9 copies/cell, respectively, by repeated integration. This integration system is a promising approach for the easy construction of yeast strains with enhanced metabolic pathways through multicopy gene expression.
참고문헌 (19)
- Murray AW, "Pedigree analysis of plasmid segregation in yeast", Cell, 34, pp.961~970 (1983)[DOI]
- Romanos MA, "Foreign gene expression in yeast: a review", Yeast, 8, pp.423~488 (1992)[DOI]
- Lee FW, "Sequential delta-integration for the regulated insertion of cloned genes in Saccharomyces cerevisiae", Biotechnol. Prog., 13, pp.368~373 (1997)[DOI]
- Kim MD, "Enhanced production of anticoagulant hirudin in recombinant Saccharomyces cerevisiae by chromosomal <TEX>${\delta}$<", TEX>-integration, 2001, pp.85, 41~48
- Sakai A, "Integration of heterologous genes into the chromosome of Saccharomyces cerevisiae using a <TEX>${\delta}$<", TEX>-sequence of yeast retrotransposon Ty, 1990, pp.33, 302~306
- Mochizuki D, "Overexpression and secretion of cellulolytic enzymes by <TEX>${\delta}$<", TEX>-sequences mediated multicopy integration of heterologous DNA sequences into the chromosomes of Saccharomyces cerevisiae, 1994, pp.77, 468~473
- Cho KM, "<TEX>${\delta}$<", TEX>-Integration of endo:exo- glucanase and <TEX>${\beta}$<, Enzyme Microb. Technol., pp.1999, 25[DOI]
- Lopes TS, "Mechanism of high-copy-number integration of pMIRY- type vectors into the ribosomal DNA of Saccharomyces cerevisiae", Gene, 105, pp.83~90 (1991)[DOI]
- Nieto A, "Stable high-copy-number integration of Aspergillus oryzae <TEX>${\alpha}$<", TEX>-amylase cDNA in an industrial baker's yeast strain, 1999, pp.15, 459~466
- Kim YH, "A versatile and general splitting technology for generating targeted YAC subclones", Appl. Microbiol. Biotechnol., 69, pp.65~70 (2005)[DOI]
- Guldener U, "A new efficient gene disruption cassette for repeated use in budding yeast", Nucleic Acids Res., 24, pp.2519~2524 (1996)[DOI]
- Kim YH, "Chromosome XII context is important for rDNA function in yeast", Nucleic Acids Res., 34, pp.2914~2924 (2006)[DOI]
- 김연희, "복제수 증폭시스템과 염색체 분단기술을 이용한 Simultaneous YAC Manipulation-Amplification (SYMA) 시스템의 개발", 생명과학회지, 20(5), pp.789~793 (2010), 한국생명과학회
- 김연희, "Bacillus subtilis 유래 재조합 endoxylanase를 이용한 xylooligosaccharide의 최적 생산", 생명과학회지, 18(1), pp.52~57 (2008), 한국생명과학회
- 이란희, "꿀벌(Apis mellifera)의 장내 세균인 Bacillus sp. HY-20이 분비하는 Xylanase의 특성", 미생물학회지, 45(4), pp.332~338 (2009), 한국미생물학회
- Chun YC, "Molecular cloning and the nucleotide sequence of a Bacillus sp. KK-1 <TEX>${\beta}$<", TEX>-xylosidase gene, 1998, pp.8, 28~33
- Sugiyama M, "Repeated chromosome splitting targeted to delta sequences in Saccharomyces cerevisiae", J. Biosci. Bioeng., 96, pp.397~400 (2003)[DOI]
- Min-Ji Kim, "Efficient Secretory Expression of Recombinant Endoxylanase from Bacillus sp. HY-20 in Saccharomyces cerevisiae", 생명과학회지, 23(7), pp.863~868 (2013), 한국생명과학회[DOI]
- Lee FW, "Sequential delta-integration for the regulated insertion of cloned genes in Saccharomyces cerevisiae", Biotechnol. Prog., 13, pp.368~373 (1997)[DOI]