24小时热门版块排行榜

返回列表

本帖产生 1 个博学EPI ，点击这里进行查看

wuweibewin

金虫 (正式写手)

博学EPI: 14
应助: 8 (幼儿园)
金币: 673.3
散金: 195
红花: 8
帖子: 634
在线: 42.8小时
虫号: 738179
注册: 2009-04-02
性别: GG
专业: 作物分子育种

[求助] Screening for Gene Regulation Mutants by Bioluminescence Imaging

--------------------------------------------------------------------------------

https://stke.sciencemag.org/cgi/ ... trans;2002/140/pl10

Sci. STKE, 9 July 2002
Vol. 2002, Issue 140, p. pl10
[DOI: 10.1126/stke.2002.140.pl10]

PROTOCOLS
Screening for Gene Regulation Mutants by Bioluminescence Imaging
Viswanathan Chinnusamy, Becky Stevenson, Byeong-ha Lee, and Jian-Kang Zhu*

Department of Plant Sciences, University of Arizona, Tucson, AZ 85721, USA.

Abstract: Because plants cannot move, they have evolved complex sensing and response systems to cope with the physical environment. Adverse environmental conditions, such as those causing abiotic stress, often cause significant losses in crop productivity and quality. Because of a paucity of well-defined visible phenotypes, conventional genetic screens have not been very successful in isolating abiotic stress signal transduction mutants of plants. Here, we describe a reporter gene-based strategy to screen for mutants affected in abiotic stress-regulated gene transcription. Our genetic screen uses the firefly luciferase reporter gene driven by the cold, drought, salt, and abscisic acid (ABA)-responsive RD29A promoter (RD29A::LUC). Arabidopsis plants transformed with the RD29A::LUC reporter emit bioluminescence in response to cold, drought, salt, or ABA treatment. After mutagenesis of these plants with ethyl methanesulfonate (EMS), mutants can be screened from the M2 population by monitoring the level of stress-inducible bioluminescence with a high-throughput, low-light imaging system. This protocol describes in detail the procedures for this luciferase reporter-based genetic screen for Arabidopsis mutants defective in abiotic stress signaling.

回复此楼