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小鼠表皮型脂肪酸结合蛋白基因(Fabpe)的克隆与染色体定位

Cloning and chromosomal localisation of the murine epidermal-type fatty acid binding protein gene (Fabpe).

作者信息

Bleck B, Hohoff C, Binas B, Rüstow B, Dixkens C, Hameister H, Börchers T, Spener F

机构信息

Department of Biochemistry, University of Münster, Wilhelm-Klemm-Str. 2, D-48149, Münster, Germany.

出版信息

Gene. 1998 Jul 17;215(1):123-30. doi: 10.1016/s0378-1119(98)00262-5.

Abstract

We succeeded in cloning the gene encoding the murine epidermal-type fatty acid binding protein (E-FABP). To avoid the screening of pseudogenes, the presence of which was shown by PCR, we designed an intron-specific probe and screened a bacterial artificial chromosome library from mouse embryonic stem cells. One of the clones obtained was analysed by restriction with various enzymes and an 11-kb EcoRI fragment with the complete gene was subcloned. The gene revealed the canonical exon/intron FABP structure consisting of four exons (112, 173, 102 and 544bp, respectively) and three introns (2217, 327 and 546bp, respectively). The exon sequences were identical with the cDNA encoding mouse E-FABP (Krieg, P., Feil, S., Fürstenberger, G., Bowden, T.G., 1993. Tumor-specific overexpression of a novel keratinocyte lipid-binding protein. Identification and characterisation of a cloned sequence activated during multistage carcinogenesis in mouse skin. J. Biol. Chem. 268, 17362-17369). Of the 5' region, 2470bp were sequenced and searched for transcription factor binding sites. Putative responsive elements within the promoter region were identified that may be responsible for the wide expression observed for E-FABP in mouse tissues. The 11-kb EcoRI fragment was used to localise Fabpe on chromosome 3 in the region 3A1-3 by fluorescence in-situ hybridisation.

摘要

我们成功克隆了编码小鼠表皮型脂肪酸结合蛋白(E-FABP)的基因。为避免筛选假基因(PCR结果显示存在假基因),我们设计了一个内含子特异性探针,并从小鼠胚胎干细胞中筛选细菌人工染色体文库。对获得的其中一个克隆进行了多种酶切分析,包含完整基因的11kb EcoRI片段被亚克隆。该基因呈现出典型的外显子/内含子FABP结构,由四个外显子(分别为112、173、102和544bp)和三个内含子(分别为2217、327和546bp)组成。外显子序列与编码小鼠E-FABP的cDNA相同(Krieg, P., Feil, S., Fürstenberger, G., Bowden, T.G., 1993. 一种新型角质形成细胞脂质结合蛋白的肿瘤特异性过表达。小鼠皮肤多阶段致癌过程中激活的克隆序列的鉴定与表征。《生物化学杂志》268, 17362 - 17369)。对5'区域的2470bp进行了测序,并搜索转录因子结合位点。在启动子区域鉴定出了可能负责E-FABP在小鼠组织中广泛表达的假定反应元件。利用11kb EcoRI片段通过荧光原位杂交将Fabpe定位到3号染色体的3A1 - 3区域。

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