Isolation of a ubiquitin-like (UBL5) gene from a screen identifying highly expressed and conserved iris genes
Isolation of a ubiquitin-like (UBL5) gene from a screen identifying highly expressed and conserved iris genes
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DOI：
10.1006/geno.2000.6439
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发表时间：
2001-01-15
期刊：
影响因子：
4.4
通讯作者：
Walter, MA
中科院分区：
文献类型：
--
作者：
Friedmann, JS;Koop, BF;Walter, MA
我们已经筛选了一个成人虹膜cDNA文库，以确定在人类和猪之间高度表达和保守的基因。我们鉴定了与猪脉络膜环cDNA探针在高严格条件下杂交的人虹膜cDNA。在检测的1568个人虹膜cDNA中，发现176个在猪脉络膜环中有高表达。176个克隆中的一个被鉴定为以前未表征的cDNA，我们将其命名为Ubiquitin-like 5基因（UBL 5）。UBL 5基因位于染色体19p13.2，其基因组结构已被检查。在染色体17p11.2上存在URL 5假基因。我们还在拟南芥、秀丽隐杆线虫、粟酒裂殖酵母和酿酒酵母中发现了UBL 5基因的同源物。泛素样基因5的北方印迹分析揭示了在每个测试组织中的表达，其中在心脏、骨骼肌、肾、肝、虹膜和淋巴母细胞中RNA表达水平最高。COS-7细胞内定位实验表明，重组UBL 5蛋白定位于细胞质内。Western分析表明，重组UBL 5蛋白约为9 kDa，从cDNA预测。使用PROTDIST程序比较UBL 5及其同源物与其他泛素样蛋白和泛素，表明URL 5基因是一类单独的泛素样基因。UBL 5基因的进一步表征将确定编码蛋白的功能以及它是否是眼部疾病的候选者。(C)北京：科学出版社.
We have screened a human adult iris cDNA library to identify genes that are highly expressed and conserved between humans and pigs. We identified human iris cDNAs that hybridized at high stringency to a porcine choroidal ring cDNA probe. Of 1568 human iris cDNAs examined, 176 were found to have high expression in porcine choroidal rings. One of the 176 clones was identified as a previously uncharacterized cDNA that we have named the Ubiquitin-like 5 gene (UBL5). The UBL5 gene is located on chromosome 19p13.2, and its genomic structure has been examined. There is a URL5 pseudogene on chromosome 17p11.2. We have also found homologues to the UBL5 gene in Arabidopsis thaliana, Caenorhabditis elegans, Schizo-saccharomyces pombe, and Saccharomyces cerevisiae. Northern blot analysis of the Ubiquitin-like gene 5 revealed expression in every tissue tested, with the highest levels of RNA expression in heart, skeletal muscle, kidney, liver, iris, and lymphoblasts. Intracellular localization experiments in COS-7 cells showed that the recombinant UBL5 protein is cytoplasmic. Western analysis demonstrated that the recombinant UBL5 protein is approximately 9 kDa, as predicted from the cDNA. A comparison between UBL5 and its homologues with other Ubiquitin-like proteins and Ubiquitin, using the PROTDIST program, suggests that the URL5 genes are a separate class of Ubiquitin-like genes. Further characterization of the UBL5 gene will determine the function of the encoded protein and whether it is a candidate for ocular disease. (C) 2001 Academic Press.