Light-mediated control of DNA transcription in yeast

被引:65
|
作者
Hughes, Robert M. [1 ]
Bolger, Steven [2 ]
Tapadia, Hersh [2 ]
Tucker, Chandra L. [1 ,3 ]
机构
[1] Duke Univ, Dept Biol, Durham, NC USA
[2] Duke Univ, Dept Biomed Engn, Durham, NC 27706 USA
[3] Univ Colorado Denver, Dept Pharmacol, Aurora, CO 80045 USA
基金
美国国家卫生研究院;
关键词
Photoreceptor; Cryptochrome; Phytochrome; Optogenetics; Two-hybrid; DNA transcription; PROTEIN-PROTEIN INTERACTIONS; SACCHAROMYCES-CEREVISIAE; GENE-EXPRESSION; GRATUITOUS INDUCTION; NUCLEOTIDE-SEQUENCE; MAMMALIAN-CELLS; PROMOTER SYSTEM; MET25; GENE; BINDING; GAL4-VP16;
D O I
10.1016/j.ymeth.2012.08.004
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
A variety of methods exist for inducible control of DNA transcription in yeast. These include the use of native yeast promoters or regulatory elements that are responsive to small molecules such as galactose, methionine, and copper, or engineered systems that allow regulation by orthogonal small molecules such as estrogen. While chemically regulated systems are easy to use and can yield high levels of protein expression, they often provide imprecise control over protein levels. Moreover, chemically regulated systems can affect many other proteins and pathways in yeast, activating signaling pathways or physiological responses. Here, we describe several methods for light mediated control of DNA transcription in vivo in yeast. We describe methodology for using a red light and phytochrome dependent system to induce transcription of genes under GAL1 promoter control, as well as blue light/cryptochrome dependent systems to control transcription of genes under GAL1 promoter or LexA operator control. Light is dose dependent, inexpensive to apply, easily delivered, and does not interfere with cellular pathways, and thus has significant advantages over chemical systems. (c) 2012 Elsevier Inc. All rights reserved.
引用
收藏
页码:385 / 391
页数:7
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