Southern Research Institute, Birmingham, AL 35205, USA.
BMC Genomics. 2010 Apr 19;11:251. doi: 10.1186/1471-2164-11-251.
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.
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.
The tools developed in this study allow for functional genomic analysis and make further development of reagents for the ferret model possible.
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。尽管它对生物医学研究很重要,但用于分子和免疫分析的试剂数量有限。我们在此介绍了一项平行测序工作,该工作从雪貂血液、肝脏、肺、脾脏和大脑的 mRNA 制成的标准化雪貂 cDNA 文库中产生了广泛的 EST(表达序列标签)数据集。
我们生成了超过 500000 条序列读段,这些序列读段组装成 16000 个部分雪貂基因。这些基因与 GenBank 中现有的雪貂序列结合,开发了一种雪貂特异性微阵列平台。使用该阵列,我们检测到了组织特异性表达模式,这些模式通过定量实时 PCR 检测得到了证实。我们还提出了一组 41 个雪貂基因,这些基因在测试的组织中具有均匀的转录谱,表明它们可用作管家基因。
本研究中开发的工具允许进行功能基因组分析,并使雪貂模型的试剂进一步开发成为可能。