Large-scale isolation of ESTs from medaka embryos and its application to medaka developmental genetics
Large-scale isolation of ESTs from medaka embryos and its application to medaka developmental genetics
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DOI：
10.1016/j.mod.2004.03.033
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发表时间：
2004-07-01
影响因子：
2.6
通讯作者：
Takeda, H
中科院分区：
文献类型：
--
作者：
Kimura, T;Jindo, T;Takeda, H
青鳉正在成为脊椎动物早期发育和器官发生研究的一种有吸引力的模式生物，日本的几个研究小组正在进行旨在产生发育缺陷突变体的大规模诱变项目。为了加强青鳉发育遗传学的研究，我们进行了大规模的ESTs分离青鳉胚胎和开发工具，方便突变分析。在这项研究中，我们从青鳉胚胎发育的不同阶段产生的文库中克隆插入cDNA的两端总共鉴定了132，082个序列。用3-素数序列进行聚类分析，最终确定了总共12，429个聚类。作为试点分析，924集群进行原位杂交，以确定其转录的空间定位。使用本研究中产生的EST序列数据，构建了具有8，091个unigenes的60-mer寡核苷酸微阵列（Medaka Microarray 8 K），并测试其在表达谱中的有用性。此外，我们已经开发了一个快速和可靠的突变作图系统，使用一套映射EST标记（M-marker 2003），涵盖了整个青鳉基因组。这些资源将加速青鳉突变分析，并为青鳉基因组计划做出重要贡献。(C)2004爱思唯尔爱尔兰有限公司保留所有权利。
The medaka is becoming an attractive model organism for the study of vertebrate early development and organogenesis and large-scale mutagenesis projects that are aimed at creating developmentally defective mutants are now being conducted by several groups in Japan. To strengthen the study of medaka developmental genetics, we have conducted a large-scale isolation of ESTs from medaka embryos and developed tools that facilitate mutant analysis. In this study, we have characterized a total of 132,082 sequences from both ends of cloned insert cDNAs from libraries generated at different stages of medaka embryo development. Clustering analysis with 3-prime sequences finally identified a total of 12,429 clusters. As a pilot analysis, 924 clusters were subjected to in situ hybridization to determine the spatial localization of their transcripts. Using EST sequence data generated in the present Study, a 60-mer oligonucleotide microarray with 8,091 unigenes (Medaka Microarray 8K) was constructed and tested for its usefulness in expression profiling. Furthermore, we have developed a rapid and reliable mutant mapping system using a set of mapped EST markers (M-marker 2003) that covers the entire medaka genome. These resources will accelerate medaka mutant analyses and make an important contribution to the medaka genome project. (C) 2004 Elsevier Ireland Ltd. All rights reserved.