Parkinson John, Blaxter Mark
Molecular Structure and Function, Hospital for Sick Children, Toronto, ON, Canada.
Methods Mol Biol. 2009;533:1-12. doi: 10.1007/978-1-60327-136-3_1.
Expressed sequence tags (ESTs) are fragments of mRNA sequences derived through single sequencing reactions performed on randomly selected clones from cDNA libraries. To date, over 45 million ESTs have been generated from over 1400 different species of eukaryotes. For the most part, EST projects are used to either complement existing genome projects or serve as low-cost alternatives for purposes of gene discovery. However, with improvements in accuracy and coverage, they are beginning to find application in fields such as phylogenetics, transcript profiling and proteomics. This volume provides practical details on the generation and analysis of ESTs. Chapters are presented which cover creation of cDNA libraries; generation and processing of sequence data; bioinformatics analysis of ESTs; and their application to phylogenetics and transcript profiling.
表达序列标签（ESTs）是通过对从cDNA文库中随机选择的克隆进行单测序反应而获得的mRNA序列片段。迄今为止，已从1400多种不同的真核生物物种中产生了超过4500万个EST。在很大程度上，EST项目用于补充现有的基因组项目，或作为基因发现目的的低成本替代方案。然而，随着准确性和覆盖范围的提高，它们开始在系统发育学、转录谱分析和蛋白质组学等领域得到应用。本书提供了关于EST生成和分析的实用细节。书中各章涵盖了cDNA文库的创建；序列数据的生成和处理；EST的生物信息学分析；以及它们在系统发育学和转录谱分析中的应用。
