翻译延伸因子2 (EF-2)在斑马鱼中的克隆与表达。

Shu-Hong Zhang, Ji-Hua Yao, Huai-Dong Song, Lu Wang, Jing-Lun Xue
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摘要

我们已经在斑马鱼中发现了一个发育调控基因翻译延伸因子2 (EF-2)。AAQ91234)。对斑马鱼EF-2的DNA序列分析表明,全长2826 bp的cDNA横跨55 ~ 2631核苷酸的开放阅读框,编码858个氨基酸的蛋白质。该基因与人类、小鼠、仓鼠、鹅、秀丽隐杆线虫和果蝇EF-2的氨基酸序列同源度分别为92、93、93、92、79和80%。斑马鱼EF-2蛋白有16个保守结构域,gtp结合结构域位于NH2端,adp核糖基化结构域位于COOH端。斑马鱼胚胎的全原位杂交表明,EF-2基因的转录本在斑马鱼胚胎发育早期就存在,并从5-体期到突口期不断发生变化。它在5-somite阶段强烈表达于整个包膜。然后染色的细胞集中在眼睛、大脑和肌肉组织中。从prim-25期开始,染色细胞仅在晶状体和小脑前部强烈出现。
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Cloning and expression of translation elongation factor 2 (EF-2) in zebrafish.

We have identified a developmentally regulated gene translation elongation factor 2 (EF-2) in zebrafish (GenBank Accession No. AAQ91234). Analysis of DNA sequence of zebrafish EF-2 shows that the 2826 bp cDNA spans an open reading frame from nucleotide 55 to 2631 and encodes a protein of 858 amino acids. It shares an identity of 92, 93, 93, 92, 79 and 80% in amino acid sequence to human, mouse, Chinese hamster, Gallus gullus, C. elegans and Drosophila EF-2, respectively. Zebrafish EF-2 protein has 16 conserved domains, GTP-binding domain is found in the NH2 terminus, and the ADP-ribosylation domain locates at the COOH terminus. Whole mount in situ hybridization on zebrafish embryos shows that the transcripts of EF-2 gene are detected during the early development of zebrafish embryo and constantly change from 5-somite stage to protruding-mouth stage. It expresses strongly throughout envelope at 5-somite stage. Then the stained cells concentrate strongly in the eyes, brain and muscle tissue. From prim-25 stage the stained cells only appear strongly in the lens and the anterior portion of the cerebellum.

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