Transcriptome sequencing and development of an expression microarray platform for the domestic ferret
Transcriptome sequencing and development of an expression microarray platform for the domestic ferret
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DOI：
10.1186/1471-2164-11-251
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发表时间：
2010-04-19
期刊：
影响因子：
4.4
通讯作者：
Jonsson, Colleen B.
中科院分区：
文献类型：
--
作者：
Bruder, Carl E.;Yao, Suxia;Jonsson, Colleen B.
背景：雪貂（Mustela putorius Furo）是研究呼吸道疾病（包括流感）的一种有吸引力的动物模型。尽管其对生物医学研究很重要，但用于分子和免疫学分析的试剂数量却受到限制。我们在这里展示了一项并行测序工作，旨在产生一个广泛的 EST（表达序列标签）数据集，该数据集源自标准化雪貂 cDNA 文库，该文库由来自雪貂血液、肝脏、肺、脾和脑的 mRNA 制成。结果：我们产生了超过 500000 个序列读数，这些序列被组装成 16000 个部分雪貂基因。这些基因与 GenBank 中可用的雪貂序列相结合，开发了雪貂特异性微阵列平台。使用该阵列，我们检测了组织特异性表达模式，并通过定量实时 PCR 测定进行了确认。我们还展示了一组 41 个雪貂基因，在测试的组织中具有均匀的转录谱，表明它们作为管家基因的有用性。结论：本研究中开发的工具允许进行功能基因组分析，并使雪貂模型试剂的进一步开发成为可能。
Background: The ferret (Mustela putorius furo) represents an attractive animal model for the study of respiratory diseases, including influenza. Despite its importance for biomedical research, the number of reagents for molecular and immunological analysis is restricted. We present here a parallel sequencing effort to produce an extensive EST (expressed sequence tags) dataset derived from a normalized ferret cDNA library made from mRNA from ferret blood, liver, lung, spleen and brain.Results: We produced more than 500000 sequence reads that were assembled into 16000 partial ferret genes. These genes were combined with the available ferret sequences in the GenBank to develop a ferret specific microarray platform. Using this array, we detected tissue specific expression patterns which were confirmed by quantitative real time PCR assays. We also present a set of 41 ferret genes with even transcription profiles across the tested tissues, indicating their usefulness as housekeeping genes.Conclusion: The tools developed in this study allow for functional genomic analysis and make further development of reagents for the ferret model possible.