Zebrafish (Danio rerio) p53 tumor suppressor gene: cDNA sequence and expression during embryogenesis.

R Cheng, B L Ford, P E O'Neal, C Z Mathews, C S Bradford, T Thongtan, D W Barnes, J D Hendricks, G S Bailey
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Abstract

Three methods were used in succession to screen a whole adult zebrafish cDNA library for expressed p53-like genes. The sequences of the resultant clones describe an open reading frame 1122 nucleotides in length, with another 43 and 940 bases of 5' and 3' untranslated sequence, respectively. The deduced amino acid sequence of the zebrafish p53 protein is 63% identical to that of trout and 48% identical to that of human p53. Two of the three zebrafish clones overlap to span the entire reported cDNA sequence and are identical in their deduced amino acid sequence over their coincident length. The third clone contains a conservative amino acid change, as well as an inserted amino acid subsequently found to be at the junction of exons 2 and 3, suggestive of alternative splicing in the p53 mRNA for this species. Northern analysis demonstrated a zebrafish p53-related transcript to be present and most abundant in zygotes and early-cleavage embryos less than 1 hour after fertilization, thereafter declining to barely detectable levels at 48 hours. A similar temporal expression was detected for the zebrafish L-myc, known to be present in maternally derived RNA, whereas zebrafish N-myc and the zebrafish homologue of the murine T gene were not detectable prior to the onset of zygotic transcription.

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斑马鱼(Danio rerio) p53肿瘤抑制基因:cDNA序列及胚胎发生过程中的表达。
用三种方法连续筛选整个成年斑马鱼cDNA文库中表达的p53样基因。得到的克隆序列描述了一个长度为1122个核苷酸的开放阅读框,另外还有43个碱基的5'和940个碱基的3'未翻译序列。推导出的斑马鱼p53蛋白的氨基酸序列与鳟鱼的相同度为63%,与人类p53相同度为48%。三个斑马鱼克隆中的两个重叠以跨越整个报道的cDNA序列,并且在其相同的长度上推导出相同的氨基酸序列。第三个克隆包含一个保守的氨基酸变化,以及随后在外显子2和3的连接处发现的插入氨基酸,这表明该物种的p53 mRNA中存在选择性剪接。Northern分析表明,斑马鱼p53相关转录物在受精后不到1小时的受精卵和早期卵裂胚胎中最丰富,此后在48小时内几乎检测不到。在斑马鱼的L-myc中检测到类似的时间表达,已知存在于母源性RNA中,而斑马鱼的N-myc和小鼠T基因的斑马鱼同源物在受精卵转录开始之前未被检测到。
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