Ishikawa K, Nagase T, Suyama M, Miyajima N, Tanaka A, Kotani H, Nomura N, Ohara O
Kazusa DNA Research Institute, Kisarazu, Chiba, Japan.
DNA Res. 1998 Jun 30;5(3):169-76. doi: 10.1093/dnares/5.3.169.
As an extension of our cDNA analysis for deducing the coding sequences of unidentified human genes, we have newly determined the sequences of 100 cDNA clones from a set of size-fractionated human brain cDNA libraries, and predicted the coding sequences of the corresponding genes, named KIAA0611 to KIAA0710. In vitro transcription-coupled translation assay was applied as the first screening to select cDNA clones which produce proteins with apparent molecular mass of 50 kDa and over. One hundred unidentified cDNA clones thus selected were then subjected to sequencing of entire inserts. The average size of the inserts and corresponding open reading frames was 4.9 kb and 2.8 kb (922 amino acid residues), respectively. Computer search of the sequences against the public databases indicated that predicted coding sequences of 87 genes were similar to those of known genes, 62% of which (54 genes) were categorized as proteins related to cell signaling/communication, cell structure/motility and nucleic acid management. The expression profiles in 10 human tissues of all the clones characterized in this study were examined by reverse transcription-coupled polymerase chain reaction and the chromosomal locations of the clones were determined by using human-rodent hybrid panels.
作为我们用于推导未鉴定人类基因编码序列的cDNA分析的扩展,我们从一组大小分级的人类脑cDNA文库中新测定了100个cDNA克隆的序列,并预测了相应基因的编码序列,命名为KIAA0611至KIAA0710。体外转录偶联翻译分析作为首次筛选方法,用于选择能产生表观分子量在50 kDa及以上蛋白质的cDNA克隆。然后对如此筛选出的100个未鉴定cDNA克隆进行全长插入片段测序。插入片段和相应开放阅读框的平均大小分别为4.9 kb和2.8 kb(922个氨基酸残基)。将这些序列与公共数据库进行计算机搜索表明,87个基因的预测编码序列与已知基因相似,其中62%(54个基因)被归类为与细胞信号传导/通讯、细胞结构/运动及核酸管理相关的蛋白质。通过逆转录偶联聚合酶链反应检测了本研究中鉴定的所有克隆在10种人类组织中的表达谱,并使用人-啮齿动物杂交板确定了这些克隆的染色体定位。