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用于粟酒裂殖酵母的FLAG和组氨酸标签载体的构建。

Construction of FLAG and histidine tagging vectors for Schizosaccharomyces pombe.

作者信息

Huang Y, Hamada M, Patel J, Maraia R J

机构信息

Laboratory of Molecular Growth Regulation, National Institute of Child Health and Human Development/NIH, Building 6, Room 3403, 6 Center Drive MSC 2753, Bethesda, MD 20892-2753, USA.

出版信息

Yeast. 2001 Mar 30;18(5):463-8. doi: 10.1002/yea.692.

Abstract

Schizosaccharomyces pombe is becoming an increasingly popular model system for investigating important cellular processes. To facilitate detection, purification and functional studies of Sz. pombe gene products, we constructed two tagging expression vectors for use in Sz. pombe. These vectors allow proteins to be expressed ectopically as fusion proteins with a FLAG epitope and six histidine residue tags attached to their N-terminus or C-terminus. The function and applicability of these vectors were examined and the results are shown using the N-terminal tagging vector encoding Sfc6p, a subunit of the Sz. pombe RNA polymerase III general transcription factor, TFIIIC.

摘要

粟酒裂殖酵母正日益成为用于研究重要细胞过程的热门模式系统。为便于对粟酒裂殖酵母基因产物进行检测、纯化和功能研究,我们构建了两个用于粟酒裂殖酵母的标签表达载体。这些载体可使蛋白质作为融合蛋白异位表达,其N端或C端带有一个FLAG表位和六个组氨酸残基标签。对这些载体的功能和适用性进行了检测,并使用编码粟酒裂殖酵母RNA聚合酶III通用转录因子TFIIIC的一个亚基Sfc6p的N端标签载体展示了结果。

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