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非洲爪蟾编码人类Ku70和Ku80自身抗原同源物的cDNA的分子克隆、测序及其在非洲爪蟾不同组织中的表达

Molecular cloning and sequencing of cDNAs encoding homologues of human Ku70 and Ku80 autoantigen from Xenopus and their expression in various Xenopus tissues.

作者信息

Yagura T, Sumi K

机构信息

Laboratory of Life Science, Department of Chemistry, Faculty of Science, Kwansei Gakuin University, Nishinomiya-shi, Hyogo 662-8501, Japan.

出版信息

Biochim Biophys Acta. 1999 Apr 14;1445(1):160-4. doi: 10.1016/s0167-4781(99)00028-7.

Abstract

We isolated cDNA clones encoding Ku70 and Ku80 homologues of Xenopus laevis from a cDNA library prepared from Xenopus oocytes. The nucleotide sequences of these Ku70 and Ku80 homologues have coding sequences of 1833 bp and a 611 aa protein, and 2178 bp and a 726 aa protein, respectively. The amino acid sequences deduced from the open reading frame of the Ku70 and Ku80 cDNA clones were highly homologous to those from Ku genes previously isolated, such as human (ca. 65% and ca. 62% identity, respectively) and mouse (ca. 65% and ca. 60%), and show a certain degree of homology to Drosophila (ca. 27% with Ku70), Caenorhabditis elegans (ca. 20% with Ku80) and Saccharomyces cerevisiae (ca. 23% and ca. 19%). Our detailed comparison of the predicted amino acid sequences among these species revealed the highly conserved octa-peptide LPFXXDIR common to both Xenopus Ku70 and Ku80 homologues in the region showing the high homology throughout the species tested. A Northern analysis using specific cDNA probes showed that Ku poly(A)+ mRNAs are expressed at high levels in Xenopus adult oocyte and testis.

摘要

我们从非洲爪蟾卵母细胞制备的cDNA文库中分离出编码非洲爪蟾Ku70和Ku80同源物的cDNA克隆。这些Ku70和Ku80同源物的核苷酸序列分别具有1833 bp的编码序列和一个611个氨基酸的蛋白质,以及2178 bp的编码序列和一个726个氨基酸的蛋白质。从Ku70和Ku80 cDNA克隆的开放阅读框推导的氨基酸序列与先前分离的Ku基因的氨基酸序列高度同源,如人类(分别约65%和62%的同一性)和小鼠(约65%和60%),并且与果蝇(与Ku70约27%)、秀丽隐杆线虫(与Ku80约20%)和酿酒酵母(约23%和19%)有一定程度的同源性。我们对这些物种预测的氨基酸序列进行的详细比较揭示,在所有测试物种中显示高度同源性的区域,非洲爪蟾Ku70和Ku80同源物都有高度保守的八肽LPFXXDIR。使用特异性cDNA探针的Northern分析表明,Ku poly(A)+ mRNA在非洲爪蟾成年卵母细胞和睾丸中高水平表达。

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