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犬脑表达序列标签的特征分析与辐射杂种图谱绘制

Characterization and radiation hybrid mapping of expressed sequence tags from the canine brain.

作者信息

Roberts Monica C, Hitte Christophe C, Hendrickson Julie A, Hoffmann Daniel E, Flickinger Gail H, Rutherford Mark S, Guyon Richard S, Galibert Francis S, Mickelson James R

机构信息

College of Veterinary Medicine, Department of Veterinary PathoBiology, University of Minnesota, 1988 Fitch Avenue, 295 Animal Science/Veterinary Medicine, St. Paul 55108, USA.

出版信息

Mamm Genome. 2003 Mar;14(3):203-13. doi: 10.1007/s00335-002-2207-2.

Abstract

Maps of the canine genome are now developing rapidly. Most of the markers on the current integrated canine radiation hybrid/genetic linkage/cytogenetic map are highly polymorphic microsatellite (type II) markers that are very useful for mapping disease loci. However, there is still an urgent need for the mapping of gene-based (type I) markers that are required for comparative mapping, as well as identifying candidate genes for disease loci that have been genetically mapped. We constructed an adult brain cDNA library as a resource to increase the number of gene-based markers on the canine genome map. Eighty-one percent of the 2700 sequenced expressed sequence tags (ESTs) represented unique sequences. The canine brain ESTs were compared with sequences in public databases to identify putative canine orthologs of human genes. One hundred nine of the canine ESTs were mapped on the latest canine radiation hybrid (RH) panel to determine the location of the respective canine gene. The addition of these new gene-based markers revealed three conserved segments (CS) between human and canine genomes previously detected by fluorescence in situ hybridization (FISH), but not by RH mapping. In addition, five new CS between dog and human were identified that had not been detected previously by RH mapping or FISH. This work has increased the number of gene-based markers on the canine RH map by approximately 30% and indicates the benefit to be gained by increasing the gene content of the current canine comparative map.

摘要

犬类基因组图谱目前正在迅速发展。当前整合的犬类辐射杂种/遗传连锁/细胞遗传图谱上的大多数标记都是高度多态的微卫星(II型)标记,这些标记对于绘制疾病基因座非常有用。然而,对于比较图谱所需的基于基因的(I型)标记的绘制,以及识别已通过基因定位的疾病基因座的候选基因,仍然迫切需要。我们构建了一个成年大脑cDNA文库,作为增加犬类基因组图谱上基于基因的标记数量的资源。在2700个测序的表达序列标签(EST)中,81%代表独特序列。将犬类大脑EST与公共数据库中的序列进行比较,以鉴定人类基因的推定犬类直系同源基因。将109个犬类EST定位到最新的犬类辐射杂种(RH)板上,以确定各自犬类基因的位置。这些新的基于基因的标记的添加揭示了人类和犬类基因组之间先前通过荧光原位杂交(FISH)检测到但未通过RH图谱检测到的三个保守区段(CS)。此外,还鉴定出了五个犬类和人类之间新的CS,这些CS以前通过RH图谱或FISH未检测到。这项工作使犬类RH图谱上基于基因的标记数量增加了约30%,并表明增加当前犬类比较图谱的基因含量将带来益处。

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