利用地塞米松诱导的转录激活系统pOp6/LhGR在拟南芥和其他植物中进行转基因诱导的通用方法

Q1 Agricultural and Biological Sciences Current protocols in plant biology Pub Date : 2019-03-12 DOI:10.1002/cppb.20089

Marketa Samalova, Charlotte Kirchhelle, Ian Moore

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引用次数: 10

摘要

使用化学诱导系统进行转基因表达是现代植物生物学研究的关键要求，因为它允许(1)在组成性表达时表达损害植物活力或肥力的转基因;(2)对转基因表达水平的时空控制。我们描述了严格调控和高响应的地塞米松诱导的基因表达系统pOp6/LhGR，它包括嵌合转录激活子LhGR和相应的pOp6启动子。诱导后，LhGR激活子与pOp6启动子结合，诱导目标基因的表达。我们提供了在不同发育阶段和不同植物物种中诱导转基因表达的详细方案，并讨论了地塞米松的稳定性及其类似物的使用。我们还介绍了新的、通用的、网关兼容的二进制矢量，现在可用于pOp6/LhGR系统。©2019 by John Wiley &儿子,Inc。

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Universal Methods for Transgene Induction Using the Dexamethasone-Inducible Transcription Activation System pOp6/LhGR in Arabidopsis and Other Plant Species

Use of chemically inducible systems for transgene expression is a crucial requirement for modern plant biology research, as it allows (1) expression of transgenes that compromise plant viability or fertility when constitutively expressed and (2) spatiotemporal control of transgene expression levels. We describe the stringently regulated and highly responsive dexamethasone-inducible gene expression system pOp6/LhGR, which comprises the chimeric transcription activator LhGR and the corresponding pOp6 promoter. Upon induction, the LhGR activator binds to the pOp6 promoter and induces expression of the target gene of interest. We provide detailed protocols for inducing transgene expression at different developmental stages and in different plant species and discuss dexamethasone stability and use of its analogs. We also introduce new, versatile, GATEWAY-compatible binary vectors that are now available for the pOp6/LhGR system. © 2019 by John Wiley & Sons, Inc.

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Current protocols in plant biology Agricultural and Biological Sciences-Plant Science

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期刊介绍： Sound and reproducible laboratory methods are the foundation of scientific discovery. Yet nuances that are critical for an experiment''s success are not captured in the primary literature but exist only as part of a lab''s oral tradition. Current Protocols in Plant Biology provides reproducible step-by-step instructions for protocols relevant to plant research. Furthermore, Current Protocols content is thoughtfully organized by topic for optimal usage and to maximize contextual knowledge. Quarterly issues allow Current Protocols in Plant Biology to constantly evolve to keep pace with the newest discoveries and developments. Current Protocols in Plant Biology is the comprehensive source for protocols in the multidisciplinary field of plant biology, providing an extensive range of protocols from basic to cutting edge. Coverage includes: Extraction and analysis of DNA, RNA, proteins Chromosome analysis Transcriptional analysis Protein expression Metabolites Plant enzymology Epigenetics Plant genetic transformation Mutagenesis Arabidopsis, Maize, Poplar, Rice, and Soybean, and more.