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北太平洋桡足类动物——美拟哲水蚤的功能基因组学资源:EST 数据库和生理微阵列。

Functional genomics resources for the North Atlantic copepod, Calanus finmarchicus: EST database and physiological microarray.

机构信息

Békésy Laboratory of Neurobiology, Pacific Biosciences Research Center, University of Hawaii at Manoa, Honolulu, HI 96822, USA.

出版信息

Comp Biochem Physiol Part D Genomics Proteomics. 2012 Jun;7(2):110-23. doi: 10.1016/j.cbd.2011.12.001. Epub 2011 Dec 29.

Abstract

The copepod, Calanus finmarchicus is a keystone species for the North Atlantic. Because of recent changes in the geographic distribution of this species, there are questions as to how this organism responds physiologically to environmental cues. Molecular techniques allow for examination and new understanding of these physiological changes. Here, we describe the development of a microarray for high-throughput studies of the physiological ecology of C. finmarchicus. An EST database was generated for this species using a normalized cDNA library derived from adult and sub-adult individuals. Sequence data were clustered into contigs and annotated using Blastx. Target transcripts were selected, and unique, 50 base-pair, oligomer probes were generated for 995 genes. Blast2GO processing provided detailed information on gene function. The selected targets included broad representation of biological processes, cellular components, and molecular functions. The microarray was tested in two sets of comparisons: adult females maintained at different food concentrations and field-caught sub-adults showing differences in lipid storage. Up-regulated and down-regulated transcripts were identified for both comparisons. Only a small subset of the genes up-regulated in low food individuals were also up-regulated in lipid-poor animals; no overlap was seen between the genes down-regulated in the two comparisons.

摘要

桡足类动物,长额象鼻鱼,是北大西洋的关键物种。由于该物种的地理分布最近发生了变化,人们对该生物如何对环境线索做出生理反应产生了疑问。分子技术可以用来研究和理解这些生理变化。在这里,我们描述了为长额象鼻鱼的生理生态学进行高通量研究的微阵列的开发。使用从成年和亚成年个体中衍生的归一化 cDNA 文库,为该物种生成了一个 EST 数据库。序列数据被聚类成 contigs,并使用 Blastx 进行注释。选择目标转录物,并为 995 个基因生成了独特的 50 碱基对寡核苷酸探针。Blast2GO 处理提供了有关基因功能的详细信息。所选目标包括广泛的生物学过程、细胞成分和分子功能。该微阵列在两组比较中进行了测试:在不同食物浓度下饲养的成年雌鱼和在野外捕获的显示脂质储存差异的亚成年个体。这两种比较都鉴定出上调和下调的转录本。在低食物个体中上调的基因中只有一小部分也在脂质缺乏的动物中上调;这两种比较中下调的基因之间没有重叠。

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