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首页> 外文期刊>Genomics >Application of functional genomic technologies in a mouse model of retinal degeneration.
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Application of functional genomic technologies in a mouse model of retinal degeneration.

机译：功能基因组技术在视网膜变性小鼠模型中的应用。

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Generation of tissue-specific, normalized and subtracted cDNA libraries has the potential to characterize the expression of rare transcriptional units not represented on Affymetrix GeneChips. Initial sequence analysis of our murine cDNA clone collections showed that as much as 86, 45, and 30% of clones are not represented on the Affymetrix Mu11k, MG-U74, and MG-430 chip sets, respectively. A detailed study that compared EST sequences of a subtracted library generated from mouse retina to those of MG-430 consensus sequences was undertaken, using UniGene build 124 as the common reference. A set of 1111 nonredundant transcript regions, not represented on the commercial array, was identified. These clusters were used as the primary filter for analyzing a data set produced by assaying samples from the Pde6b(rd1) mouse model of retinal degeneration on a 12,325-feature retinal cDNA microarray. QRT-PCR validated eight unique transcripts identified by microarray. Seven of the transcripts showed retina-specific expression. Full-length cloning strategies were applied to two of the ESTs. The genes discovered by this approach are the full-length mouse homologue of guanylate cyclase 2F (GUCY2F) and a carboxy-truncated splice variant of retinal S-antigen (SAG), known as regulators of the visual phototransduction G-protein-coupled receptor-mediated signaling pathway. These sequences have been assigned GenBank Accession Nos. and , respectively.
机译：组织特异性，标准化和扣除的cDNA文库的产生具有表征Affymetrix GeneChips上未代表的稀有转录单位表达的潜力。我们的鼠cDNA克隆集合的初步序列分析表明，Affymetrix Mu11k，MG-U74和MG-430芯片组分别没有代表多达86％，45％和30％的克隆。使用UniGene build 124作为通用参考，进行了一项详细研究，该研究将小鼠视网膜产生的扣除文库的EST序列与MG-430共有序列进行了比较。鉴定出一组1111个非冗余的转录本区域，这些区域在商业阵列上未显示。这些簇被用作主要过滤器，用于分析通过在12325个特征的视网膜cDNA微阵列上分析来自Pde6b（rd1）小鼠视网膜变性模型的样本而产生的数据集。 QRT-PCR验证了通过微阵列鉴定的八种独特的转录本。七个转录本显示视网膜特异性表达。全长克隆策略应用于两个EST。通过这种方法发现的基因是鸟苷酸环化酶2F（GUCY2F）的全长小鼠同源物和视网膜S抗原（SAG）的羧基截短剪接变体，被称为视觉光转导G蛋白偶联受体-介导的信号通路。这些序列已分别分配了GenBank登录号和。

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