Biochemical Approaches Including the Design and Use of Strains Expressing Epitope-Tagged Proteins

被引：11

作者：

Couvillion, Mary T. ^{[1
]}

Collins, Kathleen ^{[1
]}

机构：

[1] Univ Calif Berkeley, Dept Mol & Cell Biol, Berkeley, CA 94720 USA

来源：

TETRAHYMENA THERMOPHILA | 2012年 / 109卷

关键词：

INDUCIBLE-REPRESSIBLE PROMOTER; DEPENDENT RNA-POLYMERASE; TETRAHYMENA-THERMOPHILA; WIDE IDENTIFICATION; GENE; TRANSFORMATION; BIOGENESIS; SEQUENCE; REPLACEMENT; DICER;

D O I：

10.1016/B978-0-12-385967-9.00012-8

中图分类号：

Q2 [细胞生物学];

学科分类号：

071009 ; 090102 ;

摘要：

Epitope tagging is a powerful approach used to enable investigations of a cellular component by elucidating its localization, interaction partners, and/or activity targets. Successful tag-based affinity purification yields a mixture of the molecule of interest, associated proteins and nucleic acids, and nonspecific background proteins and nucleic acids, many of which can depend on details of the protocol for enrichment. This chapter provides guidelines and considerations for designing an affinity purification experiment, beginning with construction of a strain expressing a tagged subunit. Common biochemical methods for detecting protein, RNA, and DNA in Tetrahymena thermophila are also discussed.

引用

页码：347 / 355

页数：9

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