Efficient gene tagging in Arabidopsis thaliana using a gene trap approach
Efficient gene tagging in Arabidopsis thaliana using a gene trap approach
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DOI：
10.1073/pnas.94.23.12722
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发表时间：
1997-11-11
影响因子：
11.1
通讯作者：
Kushnir, S
中科院分区：
文献类型：
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作者：
Babiychuk, E;Fuanghthong, M;Kushnir, S
有关植物基因的大量DNA序列信息正迅速积累在公共数据库中，但为了从DNA序列到生物功能的进展，理想情况下每个基因的突变等位基因都应该是可用的，这里我们描述了一种基因陷阱结构，它允许我们高效地干扰拟南芥中转录的基因。在所使用的T-DNA载体中，编码新霉素磷酸转移酶II(NptII)的细菌报告基因的表达依赖于T-DNA整合到拟南芥基因组中的体内翻译融合的产生。对20个转基因株系的分析表明，12个株系是T-DNA插入突变体，中断的基因分析了编码核糖体蛋白(3个株系)、天冬氨酸tRNA合成酶、DNA连接酶、碱性结构域亮氨酸拉链DNA结合蛋白、ATP结合盒转运体和5个未知功能蛋白的基因，其中4个标记基因是拟南芥的新基因。这些结果表明，以nptII为报告基因的基因捕获率可高达80%，为系统的基因标记开辟了新的前景。
Large quantities of DNA sequence information about plant genes are rapidly accumulating in public databases, but to progress from DNA sequence to biological function a mutant allele for each of the genes ideally should be available, Here we describe a gene trap construct that allowed us to disrupt transcribed genes with a high efficiency in Arabidopsis thaliana. In the T-DNA vector used, the expression of a bacterial reporter gene coding for neomycin phosphotransferase II (nptII) depends on the in vivo generation of a translation fusion upon the T-DNA integration into the Arabidopsis genome, Analysis of 20 selected transgenic lines showed that 12 lines are T-DNA insertion mutants, The disrupted genes analyzed encoded ribosomal proteins (three lines), aspartate tRNA synthase, DNA ligase, basic-domain leucine zipper DNA binding protein, ATP-binding cassette transporter, and five proteins of unknown function, Four tagged genes were new for Arabidopsis. The results presented here suggest that gene trapping, using nptII as a reporter gene, can be as high as 80% and opens novel perspectives for systematic gene tagging in A. thaliana.