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T-DNA激活标签法

T-DNA activation tagging.

作者信息

Memelink Johan

机构信息

Institute of Biology, Clusius Laboratory, Leiden University, Leiden, The Netherlands.

出版信息

Methods Mol Biol. 2003;236:345-62. doi: 10.1385/1-59259-413-1:345.

Abstract

T-DNA activation tagging is a method to generate dominant mutations in plants or plant cells by random insertion of a T-DNA carrying constitutive enhancer elements, which can cause transcriptional activation of flanking plant genes. The method consists of generating a large number of transformed plants or plant cells using a specialized T-DNA construct, followed by selection for the desired phenotype. Subsequently, the activated plant gene is rescued from selected mutant transformants for further functional analysis. Since the exact procedure depends on the plant material and the selected phenotype, this chapter describes one specific example of T-DNA activation tagging of suspension-cultured cells, including, where possible, cross-references to more general applications of the technique.

摘要

T-DNA激活标签法是一种通过随机插入携带组成型增强子元件的T-DNA在植物或植物细胞中产生显性突变的方法,该增强子元件可导致侧翼植物基因的转录激活。该方法包括使用专门的T-DNA构建体产生大量转化植株或植物细胞,随后选择所需表型。随后,从选定的突变转化体中拯救出激活的植物基因用于进一步的功能分析。由于具体步骤取决于植物材料和所选表型,本章描述了悬浮培养细胞T-DNA激活标签法的一个具体实例,并在可能的情况下交叉引用该技术更广泛的应用。

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