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未分化小鼠胚胎癌细胞F9中表达基因的研究:低丰度mRNA的特性分析

A survey of genes expressed in undifferentiated mouse embryonal carcinoma F9 cells: characterization of low-abundance mRNAs.

作者信息

Nishiguchi S, Joh T, Horie K, Zou Z, Yasunaga T, Shimada K

机构信息

Department of Medical Genetics, Osaka University.

出版信息

J Biochem. 1994 Jul;116(1):128-39. doi: 10.1093/oxfordjournals.jbchem.a124485.

Abstract

As a first step to catalogue mRNAs present in mouse embryonal carcinoma F9 cells, 879 clones corresponding to low-abundance mRNAs were selected from among 2,896 randomly picked up clones of undifferentiated F9 cDNA libraries, using DNA probes complementary to poly(A)+RNAs prepared from undifferentiated F9 cells and to ones prepared from mouse fibroblast L cells. Five-hundred and eighty-two of the 879 clones were partially sequenced, and the subsequent homology search revealed that 201 corresponded to 180 known genes or known DNA sequences, which include not only housekeeping genes but also various tissue-specific genes. Interestingly, at least 24 of the 180 genes are development-related genes in mammals. Among these 24, those for midkine (growth and/or differentiation factor) and interferon-beta are reportedly up-regulated, and those for ECA39 (target for c-Myc regulation), REX-1 (zinc finger protein), and OCT-3 (POU-domain transcription factor) are down-regulated during the development of mouse embryonal carcinoma cells. Thirty-seven of the 582 clones matched the 36 previously reported unidentified ESTs (expressed sequence tags) and the remaining 344 corresponded to 329 novel ESTs. Therefore, partial sequencing of F9 cDNA clones corresponding to low-abundance mRNAs in F9 cells not only provides valuable information concerning development-related genes in mammals, but also many novel ESTs useful for studying mammalian genomes.

摘要

作为对小鼠胚胎癌细胞中存在的mRNA进行编目的第一步,从2896个随机挑选的未分化F9 cDNA文库克隆中,使用与从未分化F9细胞制备的poly(A)+RNA以及从小鼠成纤维细胞L细胞制备的poly(A)+RNA互补的DNA探针,挑选出了879个对应低丰度mRNA的克隆。对879个克隆中的582个进行了部分测序,随后的同源性搜索显示,其中201个对应于180个已知基因或已知DNA序列,这些序列不仅包括管家基因,还包括各种组织特异性基因。有趣的是,180个基因中至少有24个是哺乳动物中与发育相关的基因。在这24个基因中,据报道中期因子(生长和/或分化因子)和干扰素-β的基因上调,而ECA39(c-Myc调控靶点)、REX-1(锌指蛋白)和OCT-3(POU结构域转录因子)的基因在小鼠胚胎癌细胞发育过程中下调。582个克隆中的37个与36个先前报道的未鉴定EST(表达序列标签)匹配,其余344个对应于329个新的EST。因此,对F9细胞中对应低丰度mRNA的F9 cDNA克隆进行部分测序,不仅提供了有关哺乳动物发育相关基因的有价值信息,还提供了许多对研究哺乳动物基因组有用的新EST。

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