Identification and cloning of neuroblastoma-specific and nerve tissue-specific genes through compiled expression profiles.
Identification and cloning of neuroblastoma-specific and nerve tissue-specific genes through compiled expression profiles.
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DOI：
10.1093/dnares/3.5.311
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发表时间：
1996-10-31
期刊：
影响因子：
--
通讯作者：
Matsubara, K
中科院分区：
文献类型：
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作者：
Yokoyama, M;Nishi, Y;Matsubara, K
从人神经母细胞瘤细胞系CHP134的3‘-DNA文库中收集了1222个部分序列，获得了活性基因的表达谱。通过将这个表达谱与编辑的多个组织的谱进行比较，鉴定了几个仅在神经母细胞瘤细胞系的谱中出现的新基因转录本。Northern blotting进一步分析发现，在所研究的大多数人神经母细胞瘤中有两个特定的cdna克隆，在一些人神经母细胞瘤细胞系中以及在成人脑中也有三个。以这5个部分cDNAs为探针，克隆了全长的cDNA，并进行了序列测定。数据库搜索显示，它们都是新的和独特的序列：一个与细胞骨架蛋白有一些氨基酸序列相似性，两个克隆可能是转录因子，一个具有特征钾通道特性的克隆，以及一个与任何一种已知蛋白质不同源的克隆。因此，我们认为，收集3‘-定向的cdna序列，结合编辑的活性基因在多个组织中的表达谱，是发现在特定细胞或组织中特异表达的新基因的有力工具，在这种情况下，神经母细胞瘤和/或神经组织。
An expression profile of active genes in a human neuroblastoma cell line CHP134 was obtained by collecting 1222 partial sequences from a 3'-directed cDNA library representing a non-biased mRNA population. By comparing this expression profile with the compiled profiles of multiple tissues, several novel gene transcripts that appeared only in the profile of the neuroblastoma cell line were identified. Further analyses by Northern blotting revealed two specific cDNA clones that are expressed in most of the human neuroblastomas examined, and three that are in some of the human neuroblastoma cell lines as well as in the adult human brain. Full-size cDNAs were cloned using these five partial cDNA sequences as probes and sequenced. A database search revealed that they are all novel and unique sequences: one sharing some amino acid sequence similarities with a cytoskeletal protein, two clones likely to be transcriptional factors, a clone that has characteristic potassium channel properties, and a clone that is non-homologous to any one of the known proteins. Thus, we argue that the collection of 3'-directed cDNA sequences in combination with the compiled expression profiles of active genes in multiple tissues is a powerful tool for discovering novel genes that are specifically expressed in a given cell or tissue, in this case neuroblastomas and/or nerve tissue.