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叶绿体功能数据库:一个大规模的拟南芥 Ds/Spm-或 T-DNA 标签纯合系的集合,这些纯合系用于核编码的叶绿体蛋白,并对其进行系统表型分析。

The Chloroplast Function Database: a large-scale collection of Arabidopsis Ds/Spm- or T-DNA-tagged homozygous lines for nuclear-encoded chloroplast proteins, and their systematic phenotype analysis.

机构信息

RIKEN Plant Science Center, Yokohama, Kanagawa 230-0045, Japan.

出版信息

Plant J. 2010 Feb 1;61(3):529-42. doi: 10.1111/j.1365-313X.2009.04074.x. Epub 2009 Nov 14.

Abstract

A majority of the proteins of the chloroplast are encoded by the nuclear genome, and are post-translationally targeted to the chloroplast. From databases of tagged insertion lines at international seed stock centers and our own stock, we selected 3246 Ds/Spm (dissociator/suppressor-mutator) transposon- or T-DNA-tagged Arabidopsis lines for genes encoding 1369 chloroplast proteins (about 66% of the 2090 predicted chloroplast proteins) in which insertions disrupt the protein-coding regions. We systematically observed 3-week-old seedlings grown on agar plates, identified mutants with abnormal phenotypes and collected homozygous lines with wild-type phenotypes. We also identified insertion lines for which no homozygous plants were obtained. To date, we have identified 111 lines with reproducible seedling phenotypes, 122 lines for which we could not obtain homozygotes and 1290 homozygous lines without a visible phenotype. The Chloroplast Function Database presents the molecular and phenotypic information obtained from this resource. The database provides tools for searching for mutant lines using Arabidopsis Genome Initiative (AGI) locus numbers, tagged line numbers and phenotypes, and provides rapid access to detailed information on the tagged line resources. Moreover, our collection of insertion homozygotes provides a powerful tool to accelerate the functional analysis of nuclear-encoded chloroplast proteins in Arabidopsis. The Chloroplast Function Database is freely available at http://rarge.psc.riken.jp/chloroplast/. The homozygous lines generated in this project are also available from the various Arabidopsis stock centers. We have donated the insertion homozygotes to their originating seed stock centers.

摘要

叶绿体中的大多数蛋白质由核基因组编码,并经翻译后靶向到叶绿体。我们从国际种子库的标记插入系数据库和我们自己的库存中选择了 3246 个 Ds/Spm(解离剂/抑制-突变体)转座子或 T-DNA 标记的拟南芥系,用于编码 1369 种叶绿体蛋白(约 2090 种预测的叶绿体蛋白的 66%),其中插入破坏了蛋白质编码区。我们系统地观察了在琼脂平板上生长的 3 周龄幼苗,鉴定出具有异常表型的突变体,并收集了具有野生型表型的纯合系。我们还鉴定了没有获得纯合系的插入系。迄今为止,我们已经鉴定出 111 条具有可重复的幼苗表型的系,122 条我们无法获得纯合系,以及 1290 条没有可见表型的纯合系。叶绿体功能数据库呈现了从该资源中获得的分子和表型信息。该数据库提供了使用拟南芥基因组计划(AGI)基因座编号、标记线编号和表型搜索突变体系的工具,并提供了快速访问标记线资源详细信息的途径。此外,我们收集的插入纯合系为加速拟南芥核编码叶绿体蛋白的功能分析提供了有力工具。叶绿体功能数据库可在 http://rarge.psc.riken.jp/chloroplast/ 免费获取。该项目生成的纯合系也可从各种拟南芥库存中心获得。我们已将插入纯合系捐赠给其原始种子库中心。

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