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非洲爪蟾早期发育过程中表达的延伸因子1-α(EF-1αO)编码基因的分离与鉴定。

Isolation and characterization of the gene encoding EF-1 alpha O, an elongation factor 1-alpha expressed during early development of Xenopus laevis.

作者信息

Frydenberg J, Poulsen K, Petersen A K, Lund A, Olesen O F

机构信息

Division of Biostructural Chemistry, Aarhus University, Denmark.

出版信息

Gene. 1991 Dec 30;109(2):185-92. doi: 10.1016/0378-1119(91)90608-e.

Abstract

In Xenopus laevis, the gene encoding the elongation factor 1-alpha variant EF-1 alpha O, where O stands for oocyte, is expressed in oocytes and early embryos. A genomic library from X. laevis was screened with a cDNA probe coding for EF-1 alpha O. Two recombinant phages were isolated, one of which carries an entire EF-1 alpha O gene. This clone was characterized by restriction enzyme mapping and sequencing. Comparison of cDNA and genomic sequences revealed that EF-1 alpha O consists of seven exons spanning about 6.5 kb. The structure of the gene is very homologous to the human EF-1 alpha gene, as all locations of the splice junctions are conserved between the two genes. The sequence immediately upstream from the transcription start point (tsp) contains a CCAAT box, but does not contain either a TATA box or a Sp1-binding site. Interestingly, this sequence has a sequence homologous to the negative regulatory element from the TFIIIA promoter. A region located about 400 bp upstream from the tsp contains an additional number of possible regulatory sequence elements. The first intron contains G + C-rich elements which exist both isolated and as part of longer inverted repeats. Furthermore, one octamer and four Sp1-binding sites are found in this intron.

摘要

在非洲爪蟾中,编码延伸因子1-α变体EF-1αO(其中O代表卵母细胞)的基因在卵母细胞和早期胚胎中表达。用编码EF-1αO的cDNA探针筛选来自非洲爪蟾的基因组文库。分离出两个重组噬菌体,其中一个携带完整的EF-1αO基因。通过限制性酶切图谱分析和测序对该克隆进行了表征。cDNA和基因组序列的比较显示,EF-1αO由七个外显子组成,跨度约为6.5 kb。该基因的结构与人类EF-1α基因非常同源,因为两个基因之间剪接位点的所有位置都是保守的。转录起始点(tsp)上游紧邻的序列包含一个CCAAT框,但既不包含TATA框也不包含Sp1结合位点。有趣的是,该序列具有与TFIIIA启动子的负调控元件同源的序列。位于tsp上游约400 bp处的区域包含更多可能的调控序列元件。第一个内含子包含富含G + C的元件,这些元件既单独存在,也作为较长反向重复序列的一部分存在。此外,在这个内含子中发现了一个八聚体和四个Sp1结合位点。

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