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学科主题: 物理化学
题名: Efficient gene disruption in Saccharomyces cerevisiae using marker cassettes with long homologous arms prepared by the restriction-free cloning strategy
作者: Zhou, Yongjin J.1, 2;  Yang, Fan1, 2;  Zhang, Sufang1;  Tan, Haidong1;  Zhao, Zongbao K.1
通讯作者: 赵宗保
关键词: Disruption cassettes ;  Gene disruption ;  Restriction-free cloning ;  Disruption efficiency ;  Saccharomyces cerevisiae
刊名: WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY
发表日期: 2011-12-01
DOI: 10.1007/s11274-011-0756-9
卷: 27, 期:12, 页:2999-3003
收录类别: SCI
文章类型: Article
部门归属: 18
项目归属: 1816
产权排名: 1,1
WOS标题词: Science & Technology ;  Life Sciences & Biomedicine
类目[WOS]: Biotechnology & Applied Microbiology
摘要: Efficient gene disruption in Saccharomyces cerevisiae using marker cassettes with long homologous arms prepared by the restriction-free cloning strategy
研究领域[WOS]: Biotechnology & Applied Microbiology
英文摘要: Here we report an improved method for targeted gene disruption with high efficiency in S. cerevisiae, where the selection markers with long homologous arms are defined by the choice of the primer binding sites at the target locus and the disruption cassettes are constructed by restriction-free (RF) cloning strategy. Three genes, SAM1, IDH1 and IDH2, were disrupted with this method and the disruption efficiencies of SAM1 was improved several folds with much lower false-positive rates compared to the conventional one-step PCR-based gene disruption method. This approach for gene disruption cassettes construction with long flanking homologous arms may be readily applicable to facilitate targeted gene disruption in other non-conventional yeasts and fungi.
关键词[WOS]: DELETION ;  GENOME
语种: 英语
WOS记录号: WOS:000297594400029
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/115572
Appears in Collections:中国科学院大连化学物理研究所_期刊论文

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作者单位: 1.CAS, Dalian Inst Chem Phys, Div Biotechnol, Dalian 116023, Peoples R China
2.Chinese Acad Sci, Grad Univ, Beijing 100049, Peoples R China

Recommended Citation:
Zhou, Yongjin J.,Yang, Fan,Zhang, Sufang,et al. Efficient gene disruption in Saccharomyces cerevisiae using marker cassettes with long homologous arms prepared by the restriction-free cloning strategy[J]. WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY,2011,27(12):2999-3003.
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