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鸡基因组学资源:来自单一和多个组织cDNA文库的35407条ESTs测序与注释以及鸡基因索引的CAP3组装

Chicken genomics resource: sequencing and annotation of 35,407 ESTs from single and multiple tissue cDNA libraries and CAP3 assembly of a chicken gene index.

作者信息

Carre Wilfrid, Wang Xiaofei, Porter Tom E, Nys Yves, Tang Jianshan, Bernberg Erin, Morgan Robin, Burnside Joan, Aggrey Samuel E, Simon Jean, Cogburn Larry A

机构信息

Department of Animal and Food Sciences, University of Delaware, Newark, Delaware 19717, USA.

出版信息

Physiol Genomics. 2006 May 16;25(3):514-24. doi: 10.1152/physiolgenomics.00207.2005. Epub 2006 Mar 22.

Abstract

Its accessibility, unique evolutionary position, and recently assembled genome sequence have advanced the chicken to the forefront of comparative genomics and developmental biology research as a model organism. Several chicken expressed sequence tag (EST) projects have placed the chicken in 10th place for accrued ESTs among all organisms in GenBank. We have completed the single-pass 5'-end sequencing of 37,557 chicken cDNA clones from several single and multiple tissue cDNA libraries and have entered 35,407 EST sequences into GenBank. Our chicken EST sequences and those found in public databases (on July 1, 2004) provided a total of 517,727 public chicken ESTs and mRNAs. These sequences were used in the CAP3 assembly of a chicken gene index composed of 40,850 contigs and 79,192 unassembled singlets. The CAP3 contigs show a 96.7% match to the chicken genome sequence. The University of Delaware (UD) EST collection (43,928 clones) was assembled into 19,237 nonredundant sequences (13,495 contigs and 5,742 unassembled singlets). The UD collection contains 6,223 unique sequences that are not found in other public EST collections but show a 76% match to the chicken genome sequence. Our chicken contig and singlet sequences were annotated according to the highest BlastX and/or BlastN hits. The UD CAP3 contig assemblies and singlets are searchable by nucleotide sequence or key word (http://cogburn.dbi.udel.edu), and the cDNA clones are readily available for distribution from the chick EST website and clone repository (http://www.chickest.udel.edu). The present paper describes the construction and normalization of single and multiple tissue chicken cDNA libraries, high-throughput EST sequencing from these libraries, the CAP3 assembly of a chicken gene index from all public ESTs, and the identification of several nonredundant chicken gene sets for production of custom DNA microarrays.

摘要

鸡的易获取性、独特的进化地位以及最近组装完成的基因组序列,使其作为一种模式生物,在比较基因组学和发育生物学研究领域处于前沿位置。几个鸡表达序列标签(EST)项目已使鸡在GenBank中所有生物累积EST数量方面位居第十。我们已完成了来自多个单组织和多组织cDNA文库的37557个鸡cDNA克隆的单通道5'端测序,并将35407条EST序列录入GenBank。我们的鸡EST序列以及公共数据库(2004年7月1日)中发现的序列,共计提供了517727条公共鸡EST和mRNA。这些序列被用于构建由40850个重叠群和79192个未组装单序列组成的鸡基因索引的CAP3组装。CAP3重叠群与鸡基因组序列的匹配度为96.7%。特拉华大学(UD)的EST文库(43928个克隆)被组装成19237个非冗余序列(13495个重叠群和5742个未组装单序列)。UD文库包含6223个独特序列,这些序列在其他公共EST文库中未被发现,但与鸡基因组序列的匹配度为76%。我们的鸡重叠群和单序列根据最高的BlastX和/或BlastN比对结果进行注释。UD的CAP3重叠群组装和单序列可通过核苷酸序列或关键词进行搜索(http://cogburn.dbi.udel.edu),并且cDNA克隆可从鸡EST网站和克隆库(http://www.chickest.udel.edu)轻松获取用于分发。本文描述了单组织和多组织鸡cDNA文库的构建与标准化、这些文库的高通量EST测序、基于所有公共EST构建鸡基因索引的CAP3组装,以及鉴定几个用于生产定制DNA微阵列的非冗余鸡基因集。

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