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在构巢曲霉中快速生产基因替换构建体并生成绿色荧光蛋白标记的着丝粒标记

Rapid production of gene replacement constructs and generation of a green fluorescent protein-tagged centromeric marker in Aspergillus nidulans.

作者信息

Yang Lin, Ukil Leena, Osmani Aysha, Nahm Francis, Davies Jonathan, De Souza Colin P C, Dou Xiaowei, Perez-Balaguer Ariadna, Osmani Stephen A

机构信息

Department of Molecular Genetics, The Ohio State University, 804 Riffe Building, 496 West 12th Ave., Columbus, OH 43210, USA.

出版信息

Eukaryot Cell. 2004 Oct;3(5):1359-62. doi: 10.1128/EC.3.5.1359-1362.2004.

DOI:10.1128/EC.3.5.1359-1362.2004
PMID:15470263
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC522605/
Abstract

A method to rapidly generate gene replacement constructs by fusion PCR is described for Aspergillus nidulans. The utility of the approach is demonstrated by green fluorescent protein (GFP) tagging of A. nidulans ndc80 to visualize centromeres through the cell cycle. The methodology makes possible large-scale GFP tagging, promoter swapping, and deletion analysis of A. nidulans.

摘要

本文描述了一种通过融合PCR快速生成构巢曲霉基因置换构建体的方法。通过对构巢曲霉ndc80进行绿色荧光蛋白(GFP)标记以在细胞周期中可视化着丝粒,证明了该方法的实用性。该方法使构巢曲霉的大规模GFP标记、启动子交换和缺失分析成为可能。

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本文引用的文献

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A monomeric red fluorescent protein.一种单体红色荧光蛋白。
Proc Natl Acad Sci U S A. 2002 Jun 11;99(12):7877-82. doi: 10.1073/pnas.082243699.
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PCR-mediated generation of a gene disruption construct without the use of DNA ligase and plasmid vectors.通过聚合酶链式反应(PCR)生成基因破坏构建体,无需使用DNA连接酶和质粒载体。
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The Ndc80p complex from Saccharomyces cerevisiae contains conserved centromere components and has a function in chromosome segregation.来自酿酒酵母的Ndc80p复合物包含保守的着丝粒成分,并在染色体分离中发挥作用。
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A rapid method for efficient gene replacement in the filamentous fungus Aspergillus nidulans.一种在丝状真菌构巢曲霉中进行高效基因替换的快速方法。
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G1-phase and B-type cyclins exclude the DNA-replication factor Mcm4 from the nucleus.G1期和B型细胞周期蛋白将DNA复制因子Mcm4排除在细胞核外。
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