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8号染色体p21.3区域两个新基因的分子克隆与特性分析

Molecular cloning and characterization of two novel genes on chromosome 8p21.3.

作者信息

Kinjo T, Isomura M, Iwamasa T, Nakamura Y

机构信息

Laboratory of Molecular Medicine, University of Tokyo, Japan.

出版信息

J Hum Genet. 2000;45(1):12-7. doi: 10.1007/s100380050003.

Abstract

Through large-scale sequencing of genomic DNA from human chromosome 8p22-p21.3 we have isolated two novel genes, designated GK1 and G5. Their predicted products showed no significant similarity to any known proteins in public databases. A comparison of GK1 cDNA sequences, which encode a 1270-amino-acid protein, with corresponding genomic DNA sequences revealed that this gene consists of 15 exons and spans an approximately 113-kb genomic region. Northern blot analysis revealed ubiquitous expression of 7.0- and 4.4-kb transcripts; in addition, we detected a 5.0-kb skeletal muscle-specific transcript and a 4.0-kb transcript specifically expressed in heart and pancreas. Computer and immunocytochemical analyses of a GK1 Green fluolesent protein (GFP) fused construct indicated that the gene product, which contains putative leucine-zipper domains, was likely to be a mitochondrial protein. The other novel gene, G5, expressed four transcripts (4.2, 2.2, 1.7, and 1.0-kb) ubiquitously; the longer three transcripts, which differed only in the 3'-non coding region, encoded identical 397-amino-acid peptides. The G5 gene consists of 14 exons and spans approximately 52 kb of genomic DNA; its deduced 397-amino acid product appears to contain coiled-coil domains and a proline-rich region, and to be located in cytoplasm.

摘要

通过对人类8号染色体p22 - p21.3区域的基因组DNA进行大规模测序,我们分离出了两个新基因,命名为GK1和G5。它们的预测产物与公共数据库中任何已知蛋白质均无显著相似性。对编码1270个氨基酸蛋白质的GK1 cDNA序列与相应基因组DNA序列进行比较,发现该基因由15个外显子组成,跨越约113 kb的基因组区域。Northern印迹分析显示,存在7.0 kb和4.4 kb转录本的普遍表达;此外,我们检测到一个5.0 kb的骨骼肌特异性转录本以及一个在心脏和胰腺中特异性表达的4.0 kb转录本。对GK1绿色荧光蛋白(GFP)融合构建体进行的计算机分析和免疫细胞化学分析表明,该基因产物含有假定的亮氨酸拉链结构域,可能是一种线粒体蛋白。另一个新基因G5普遍表达四种转录本(4.2、2.2、1.7和1.0 kb);较长的三种转录本仅在3'非编码区有所不同,编码相同的397个氨基酸的肽段。G5基因由14个外显子组成,跨越约52 kb的基因组DNA;其推导的397个氨基酸产物似乎含有卷曲螺旋结构域和富含脯氨酸的区域,且定位于细胞质中。

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