EFFICIENT CLONING OF CDNAS OF RETINOIC ACID-RESPONSIVE GENES IN P19 EMBRYONAL CARCINOMA-CELLS AND CHARACTERIZATION OF A NOVEL MOUSE GENE, STRA1 (MOUSE LERK-2/EPLG2)
EFFICIENT CLONING OF CDNAS OF RETINOIC ACID-RESPONSIVE GENES IN P19 EMBRYONAL CARCINOMA-CELLS AND CHARACTERIZATION OF A NOVEL MOUSE GENE, STRA1 (MOUSE LERK-2/EPLG2)
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DOI：
10.1006/dbio.1995.1226
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发表时间：
1995-08-01
影响因子：
2.7
通讯作者：
CHAMBON, P
中科院分区：
文献类型：
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作者：
BOUILLET, P;OULADABDELGHANI, M;CHAMBON, P
多能小鼠P19胚胎癌（EC）细胞已被广泛用作发育模型系统，因为它们可以在视黄酸（RA）存在下分化成所有三个胚层的衍生物，这取决于RA剂量和培养条件。几个基因的表达已被证明是诱导RA处理的P19 EC细胞，有趣的是，这些基因中的一些可能在小鼠胚胎发生过程中发挥重要作用。鉴于越来越多的证据表明，RA是脊椎动物发育过程中的一个重要的信号分子，我们已经启动了一项研究，旨在系统分离的基因，其表达诱导P19细胞在不同的时间后，暴露于RA。在这里，我们描述了一种有效的差异消减杂交克隆策略，用于确定额外的RA反应基因在P19细胞。50个不同的cDNA片段，对应于RA诱导的基因进行了分离。10个cDNA代表已知基因，其中4个已经被描述为RA诱导的，而其余40个对应于新的基因。这些cDNA序列中的许多代表低丰度mRNA。RA治疗后的mRNA积累的动力学分析使我们能够表征四类RA响应基因。我们还报道了这些新的RA诱导基因之一Stral在小鼠胚胎和成年组织中的序列和表达模式，并表明它对应于Cek 5受体蛋白酪氨酸激酶的小鼠配体，（C）1995 Academic Press，Inc.
Pluripotent mouse P19 embryonal carcinoma (EC) cells have been extensively used as a developmental model system because they can differentiate in the presence of retinoic acid (RA) into derivatives of all three germ layers depending on RA dosage and culture conditions. The expression of several genes has been shown to be induced in RA-treated P19 EC cells and, interestingly, some of these genes may play important roles during mouse embryogenesis. In view of the increasing evidence that RA is a crucial signaling molecule during vertebrate development, we have initiated a study aimed at the systematic isolation of genes whose expression is induced in P19 cells at various times after exposure to RA. We describe here an efficient differential subtractive hybridization cloning strategy which was used to identify additional RA-responsive genes in P19 cells. Fifty different cDNA fragments corresponding to RA-induced genes were isolated. Ten cDNAs represent known genes, 4 of which have already been described as RA-inducible, while the remaining 40 correspond to novel genes. Many of these cDNA sequences represent low-abundance mRNAs. Kinetic analysis of mRNA accumulation following RA treatment allowed us to characterize four classes of RA-responsive genes. We also report the sequence and expression pattern in mouse embryos and adult tissues of one of these novel RA-inducible genes, Stral, and show that it corresponds to the mouse ligand for the Cek5 receptor protein-tyrosine kinase, (C) 1995 Academic Press, Inc.