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编码RBM家族一个新成员的人类基因的cDNA克隆、染色体定位及基因组结构

cDNA cloning, chromosome assignment, and genomic structure of a human gene encoding a novel member of the RBM family.

作者信息

Stover C, Gradl G, Jentsch I, Speicher M R, Wieser R, Schwaeble W

机构信息

Department of Microbiology and Immunology, University of Leicester, UK.

出版信息

Cytogenet Cell Genet. 2001;92(3-4):225-30. doi: 10.1159/000056908.

Abstract

We have cloned and characterised a novel human gene mapping to chromosome 20q11.2. A partial transcript was initially isolated from a human cDNA library transcribed from RNA of the colon carcinoma cell line T-84. In order to determine the full coding sequence of this novel mRNA, we isolated seven cDNA clones from a human cDNA library transcribed from RNA of the acute monocytic leukemia cell line THP1 by colony hybridization. On Northern blot analysis of four human cell lines, the cDNAs isolated hybridize with an abundantly expressed mRNA species of 3.5 kb. A full-length cDNA transcript of this novel mRNA has an open reading frame of 2,796 bp encoding a protein with a calculated molecular weight of 97 kDa. Two repetitive structural consensus motifs are contained within the deduced protein sequence, namely five distinct RNA binding motifs and two proline rich regions. The derived protein sequence also contains putative transmembrane domains. These structural motifs identify this novel protein as a member of an expanding protein family containing RNA binding motifs (RBM). As seen from recently completed sequence of the genomic area encoding this novel mRNA by the Sanger Centre Human Genome Project, the coding region of this gene, RBM12, is intronless.

摘要

我们已经克隆并鉴定了一个定位于20号染色体q11.2区域的新型人类基因。最初从结肠癌细胞系T - 84的RNA转录而成的人cDNA文库中分离出一个部分转录本。为了确定这种新型mRNA的完整编码序列,我们通过菌落杂交从急性单核细胞白血病细胞系THP1的RNA转录而成的人cDNA文库中分离出7个cDNA克隆。在对4种人类细胞系进行Northern印迹分析时,分离出的cDNA与一种大量表达的3.5 kb mRNA杂交。这种新型mRNA的全长cDNA转录本有一个2796 bp的开放阅读框,编码一种计算分子量为97 kDa的蛋白质。推导的蛋白质序列中包含两个重复的结构共有基序,即五个不同的RNA结合基序和两个富含脯氨酸的区域。推导的蛋白质序列还包含推定的跨膜结构域。这些结构基序将这种新型蛋白质鉴定为包含RNA结合基序(RBM)的不断扩大的蛋白质家族的成员。从桑格中心人类基因组计划最近完成的编码这种新型mRNA的基因组区域序列可以看出,该基因RBM12的编码区无内含子。

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