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来自斑马鱼胚胎心脏cDNA文库的表达序列标签的鉴定、表征及定位

Identification, characterization, and mapping of expressed sequence tags from an embryonic zebrafish heart cDNA library.

作者信息

Ton C, Hwang D M, Dempsey A A, Tang H C, Yoon J, Lim M, Mably J D, Fishman M C, Liew C C

机构信息

Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, Ontario M5G 1L5, Canada.

出版信息

Genome Res. 2000 Dec;10(12):1915-27. doi: 10.1101/gr.10.12.1915.

Abstract

The generation of expressed sequence tags (ESTs) has proven to be a rapid and economical approach by which to identify and characterize expressed genes. We generated 5102 ESTs from a 3-d-old embryonic zebrafish heart cDNA library. Of these, 57.6% matched to known genes, 14.2% matched only to other ESTs, and 27.8% showed no match to any ESTs or known genes. Clustering of all ESTs identified 359 unique clusters comprising 1771 ESTs, whereas the remaining 3331 ESTs did not cluster. This estimates the number of unique genes identified in the data set to be approximately 3690. A total of 1242 unique known genes were used to analyze the gene expression patterns in the zebrafish embryonic heart. These were categorized into seven categories on the basis of gene function. The largest class of genes represented those involved in gene/protein expression (25.9% of known transcripts). This class was followed by genes involved in metabolism (18.7%), cell structure/motility (16.4%), cell signaling and communication (9.6%), cell/organism defense (7.1%), and cell division (4.4%). Unclassified genes constituted the remaining 17.91%. Radiation hybrid mapping was performed for 102 ESTs and comparison of map positions between zebrafish and human identified new synteny groups. Continued comparative analysis will be useful in defining the boundaries of conserved chromosome segments between zebrafish and humans, which will facilitate the transfer of genetic information between the two organisms and improve our understanding of vertebrate evolution.

摘要

已证明表达序列标签(EST)的生成是一种快速且经济的方法,可用于鉴定和表征表达的基因。我们从一个3日龄斑马鱼胚胎心脏cDNA文库中生成了5102个EST。其中,57.6%与已知基因匹配,14.2%仅与其他EST匹配,27.8%与任何EST或已知基因均不匹配。对所有EST进行聚类分析,确定了359个独特的聚类,包含1771个EST,而其余3331个EST未聚类。由此估计数据集中鉴定出的独特基因数量约为3690个。总共1242个独特的已知基因用于分析斑马鱼胚胎心脏中的基因表达模式。根据基因功能,这些基因被分为七类。最大的一类基因是那些参与基因/蛋白质表达的基因(占已知转录本的25.9%)。其次是参与代谢的基因(18.7%)、细胞结构/运动(16.4%)、细胞信号传导和通讯(9.6%)、细胞/生物体防御(7.1%)以及细胞分裂(4.4%)。未分类的基因占其余的17.91%。对102个EST进行了辐射杂种图谱分析,斑马鱼和人类图谱位置的比较确定了新的同线群。持续的比较分析将有助于界定斑马鱼和人类之间保守染色体片段的边界,这将促进两种生物之间遗传信息的传递,并增进我们对脊椎动物进化的理解。

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