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利用萤火虫荧光素酶报告基因对拟南芥基因组进行基因捕获。

Gene trapping of the Arabidopsis genome with a firefly luciferase reporter.

作者信息

Yamamoto Yoshiharu Y, Tsuhara Yumi, Gohda Kazuhito, Suzuki Kumiko, Matsui Minami

机构信息

Plant Function Exploration Team, Plant Functional Genomics Research Group, Genomic Sciences Center, RIKEN Yokohama Institute, 1-7-22 Suyehiro-cho, Tsurumi-ku, Yokohama, Kanagawa 230-0045, Japan.

出版信息

Plant J. 2003 Jul;35(2):273-83. doi: 10.1046/j.1365-313x.2003.01797.x.

Abstract

Experiments with gene-trap vectors containing the firefly luciferase (LUC) reporter genes were carried out with the aim of analyzing functions of the Arabidopsis genome. Studies with protein fusion-type trap vectors as well as an internal ribosome entry site (IRES)-assisted non-fusion-type vector revealed that both types of vectors were suitable for gene trapping in Arabidopsis, although there were some differences in trapping efficiencies. The established trap lines were subjected to analyses for light responses, demonstrating the powerful and unique applications of a LUC-trapping system. A systematic survey of the insertion sites of the T-DNAs in LUC-expressing lines revealed 12-41% gene-trapping efficiencies depending on the vector. We demonstrate that the LUC-trapping system provides a unique system with which to monitor temporal expression of plant genes.

摘要

利用含有萤火虫荧光素酶(LUC)报告基因的基因捕获载体进行了实验,目的是分析拟南芥基因组的功能。对蛋白质融合型捕获载体以及内部核糖体进入位点(IRES)辅助的非融合型载体的研究表明,尽管捕获效率存在一些差异,但这两种类型的载体都适用于拟南芥中的基因捕获。对建立的捕获株系进行了光反应分析,证明了LUC捕获系统的强大而独特的应用。对LUC表达株系中T-DNA插入位点的系统调查显示,根据载体不同,基因捕获效率为12%-41%。我们证明,LUC捕获系统提供了一个独特的系统,可用于监测植物基因的瞬时表达。

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