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未鉴定的人类基因编码序列的预测。XX. 来自大脑的100个新cDNA克隆的完整序列,这些克隆在体外编码大蛋白。

Prediction of the coding sequences of unidentified human genes. XX. The complete sequences of 100 new cDNA clones from brain which code for large proteins in vitro.

作者信息

Nagase T, Nakayama M, Nakajima D, Kikuno R, Ohara O

机构信息

Kazusa DNA Research Institute, Kisarazu, Chiba, Japan.

出版信息

DNA Res. 2001 Apr 27;8(2):85-95. doi: 10.1093/dnares/8.2.85.

Abstract

To accumulate information on the coding sequences of unidentified genes, we have carried out a sequencing project of human cDNA clones which encode large proteins. We herein present the entire sequences of 100 cDNA clones of unidentified human genes, named KIAA1776 and KIAA1780-KIAA1878, from size-fractionated cDNA libraries derived from human fetal brain, adult whole brain, hippocampus and amygdala. Most of the cDNA clones to be entirely sequenced were selected as cDNAs which were shown to have coding potentiality by in vitro transcription/translation experiments, and some clones were chosen by using computer-assisted analysis of terminal sequences of cDNAs. Three of these clones (fibrillin2/KIAA1776, MEGF10/KIAA1780 and MEGF11/KIAA1781) were isolated as genes encoding proteins with multiple EGF-like domains by motif-trap screening. The average sizes of the inserts and corresponding open reading frames of eDNA clones analyzed here reached 4.7 kb and 2.4 kb (785 amino acid residues), respectively. From the results of homology and motif searches against the public databases, the functional categories of the predicted gene products of 54 genes were determined; 93% of these predicted gene products (50 gene products) were classified as proteins related to cell signaling/communication, nucleic acid management, or cell structure/motility. To collect additional information on these genes, their expression profiles were also studied in 10 human tissues, 8 brain regions, spinal cord, fetal brain and fetal liver by reverse transcription-coupled polymerase chain reaction, products of which were quantified by enzyme-linked immunosorbent assay.

摘要

为了积累未鉴定基因编码序列的信息,我们开展了一个人类cDNA克隆测序项目,这些克隆编码大蛋白。我们在此展示了100个未鉴定人类基因的cDNA克隆的完整序列,这些基因命名为KIAA1776以及KIAA1780 - KIAA1878,它们来自于从人类胎儿脑、成人全脑、海马体和杏仁核中分离得到的大小分级cDNA文库。大部分要进行全序列测定的cDNA克隆是通过体外转录/翻译实验被选为具有编码潜力的cDNA,还有一些克隆是通过对cDNA末端序列进行计算机辅助分析挑选出来的。其中三个克隆(原纤蛋白2/KIAA1776、多个表皮生长因子样结构域蛋白10/MEGF10和多个表皮生长因子样结构域蛋白11/MEGF11)是通过基序捕获筛选分离出来的,它们编码的蛋白质含有多个表皮生长因子样结构域。这里分析的cDNA克隆的插入片段和相应开放阅读框的平均大小分别达到了4.7 kb和2.4 kb(785个氨基酸残基)。通过对公共数据库进行同源性和基序搜索,确定了54个基因预测基因产物的功能类别;这些预测基因产物中的93%(50个基因产物)被归类为与细胞信号传导/通讯、核酸管理或细胞结构/运动相关的蛋白质。为了收集关于这些基因的更多信息,还通过逆转录偶联聚合酶链反应研究了它们在10种人类组织、8个脑区、脊髓、胎儿脑和胎儿肝脏中的表达谱,其产物通过酶联免疫吸附测定进行定量。

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