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小鼠GMFB(Gmfb)全长克隆的鉴定与分离,GMFB是一种假定的细胞内激酶调节剂,在端脑中差异表达。

Identification and isolation of a full-length clone of mouse GMFB (Gmfb), a putative intracellular kinase regulator, differentially expressed in telencephalon.

作者信息

Bourgeois F, Guimiot F, Mas C, Bulfone A, Levacher B, Moalic J M, Simonneau M

机构信息

Neurogénétique, INSERM E9935, Hôpital Robert Debré, Paris, France.

出版信息

Cytogenet Cell Genet. 2001;92(3-4):304-9. doi: 10.1159/000056919.

Abstract

We identified new transcribed sequences, using a differential display paradigm to select genes expressed in proliferating neuroblasts from mouse telencephalon at 10 days of embryonic development. In this systematic search, we isolated a 361-bp partial 3' untranslated region (3' UTR) homologous to the 3' UTR of the human gene encoding a putative intracellular kinase regulator, glia maturation factor beta (GMFB). We cloned a full-length, 4,311-bp mouse cDNA containing a 270-bp 5' UTR, a 3,615-bp 3' UTR, and an open reading frame of 426 nucleotides encoding a putative 142 amino-acid protein, identical to human GMFB, with the exception of two amino acids. This 4.3-kb transcript is present in a variety of adult tissues and is developmentally regulated as shown by Northern blot analysis. Differential expression in telencephalon was demonstrated by quantification of radioactive relative RT-PCR and confirmed by in situ hybridization. The isolation of this full-length clone of mouse Gmfb should facilitate investigation of the intracellular mechanisms involved in the development of telencephalon.

摘要

我们利用差异显示技术,从胚胎发育10天的小鼠端脑中选择在增殖神经母细胞中表达的基因,从而鉴定出新的转录序列。在这项系统研究中,我们分离出一个361 bp的部分3'非翻译区(3'UTR),它与人类基因的3'UTR同源,该人类基因编码一种假定的细胞内激酶调节剂——神经胶质成熟因子β(GMFB)。我们克隆了一个全长4311 bp的小鼠cDNA,它包含一个270 bp的5'UTR、一个3615 bp的3'UTR以及一个426个核苷酸的开放阅读框,编码一个假定的142个氨基酸的蛋白质,除了两个氨基酸外,与人GMFB相同。这种4.3 kb的转录本存在于多种成年组织中,并且如Northern印迹分析所示,其表达受发育调控。通过放射性相对RT-PCR定量证实了端脑中的差异表达,并通过原位杂交得到了确认。小鼠Gmfb全长克隆的分离应有助于研究端脑发育所涉及的细胞内机制。

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