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未鉴定的人类基因编码序列的预测。十七。来自大脑的100个新的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. XVII. The complete sequences of 100 new cDNA clones from brain which code for large proteins in vitro.

作者信息

Nagase T, Kikuno R, Ishikawa K, Hirosawa M, Ohara O

机构信息

Kazusa DNA Research Institute, Kisarazu, Chiba, Japan.

出版信息

DNA Res. 2000 Apr 28;7(2):143-50. doi: 10.1093/dnares/7.2.143.

Abstract

To provide information regarding the coding sequences of unidentified human genes, we have conducted a sequencing project of human cDNAs which encode large proteins. We herein present the entire sequences of 100 cDNA clones of unknown human genes, named KIAA1444 to KIAA1543, from two sets of size-fractionated human adult and fetal brain cDNA libraries. The average sizes of the inserts and corresponding open reading frames of cDNA clones analyzed here were 4.4 kb and 2.6 kb (856 amino acid residues), respectively. Database searches of the predicted amino acid sequences classified 53 predicted gene products into the following five functional categories: cell signaling/communication, nucleic acid management, cell structure/motility, protein management and metabolism. It was also revealed that homologues for 32 KIAA gene products were detected in the databases, which were similar in sequence through almost their entire regions. Additionally, the chromosomal loci of the genes were determined by using human-rodent hybrid panels unless their chromosomal loci were already assigned in the public databases. The expression levels of the genes were monitored in spinal cord, fetal brain and fetal liver, as well as in 10 human tissues and 8 brain regions, by reverse transcription-coupled polymerase chain reaction, products of which were quantified by enzyme-linked immunosorbent assay.

摘要

为了提供关于未鉴定的人类基因编码序列的信息,我们开展了一项人类编码大蛋白的cDNA测序计划。在此,我们展示了来自两组经大小分级的人类成人和胎儿脑cDNA文库的100个未知人类基因cDNA克隆的完整序列,命名为KIAA1444至KIAA1543。此处分析的cDNA克隆的插入片段和相应开放阅读框的平均大小分别为4.4 kb和2.6 kb(856个氨基酸残基)。对预测的氨基酸序列进行数据库搜索,将53个预测的基因产物分为以下五个功能类别:细胞信号传导/通讯、核酸管理、细胞结构/运动、蛋白质管理和代谢。还发现,在数据库中检测到32个KIAA基因产物的同源物,它们在几乎整个区域的序列都相似。此外,除非这些基因的染色体定位已在公共数据库中指定,否则通过使用人-鼠杂交板来确定这些基因的染色体定位。通过逆转录偶联聚合酶链反应监测这些基因在脊髓、胎儿脑和胎儿肝脏以及10种人类组织和8个脑区中的表达水平,其产物通过酶联免疫吸附测定进行定量。

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