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质体精氨酸tRNA-CCG基因敲除苔藓中的转录谱分析:小立碗藓质体DNA微阵列的构建

Transcript profiling in plastid arginine tRNA-CCG gene knockout moss: construction of Physcomitrella patens plastid DNA microarray.

作者信息

Nakamura T, Sugiura C, Kobayashi Y, Sugita M

机构信息

Center for Gene Research, Nagoya University, Nagoya 464-8602, Japan.

出版信息

Plant Biol (Stuttg). 2005 May;7(3):258-65. doi: 10.1055/s-2005-865620.

Abstract

The moss Physcomitrella patens is a newly established model plant that is widely used for the characterization of gene function by targeted gene knockout or over-expression. The target gene disruption occurs in both the nuclear and chloroplast genomes. We applied DNA microarray technology to the P. patens plastid genome for large-scale analysis of transcripts. A microarray was constructed containing 108 DNA fragments to detect all annotated plastid genes. We analyzed the transcript profile in a knockout transformant for the arginine tRNA gene, trnR-CCG, and confirmed previous results that rbcL and psaI transcripts accumulate in similar levels to wild-type moss, and accD transcript level is higher than those of wild-type moss. Additionally, the plastid DNA microarray revealed that most plastid genes were expressed at similar levels in wild-type and transformant mosses. This indicates that trnR-CCG is not essential for the expression of plastid genes.

摘要

小立碗藓是一种新建立的模式植物,广泛用于通过靶向基因敲除或过表达来鉴定基因功能。目标基因的破坏发生在核基因组和叶绿体基因组中。我们将DNA微阵列技术应用于小立碗藓的质体基因组,以进行转录本的大规模分析。构建了一个包含108个DNA片段的微阵列,用于检测所有注释的质体基因。我们分析了精氨酸tRNA基因trnR-CCG敲除转化体中的转录本谱,并证实了先前的结果,即rbcL和psaI转录本的积累水平与野生型苔藓相似,而accD转录本水平高于野生型苔藓。此外,质体DNA微阵列显示,大多数质体基因在野生型和转化体苔藓中的表达水平相似。这表明trnR-CCG对质体基因的表达不是必需的。

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