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玻璃海鞘RNA结合蛋白cDNA的分离与鉴定:发育过程中的时空表达

Isolation and characterization of the cDNA for a Ciona intestinalis RNA binding protein: spatial and temporal expression during development.

作者信息

Piscopo A, Branno M, Aniello F, Corrado M, Piscopo M, Fucci L

机构信息

Laboratory of Biochemistry and Molecular Biology, Stazione Zoologica Anton Dorhn, Naples, Italy.

出版信息

Differentiation. 2000 Aug;66(1):23-30. doi: 10.1046/j.1432-0436.2000.066001023.x.

Abstract

A cDNA encoding an RNA binding protein has been isolated from a cDNA library prepared from larvae of the ascidian Ciona intestinalis. The putative protein of 162 amino acids contained in the N-terminal region one copy of the consensus sequence RNA binding domain and in the C-terminal region a glycine-rich domain. The in vitro translated protein bound various RNA homopolymers, preferentially polyU, polyA, and polyG, and the binding was affected by increasing ionic strength. Northern blot analysis revealed a single transcript of about 0.7 kb in length that was present during embryonic development with two major peaks of accumulation at gastrula and larval stages. Whole-mount in situ hybridization experiments on embryos at different stages of development showed gene expression mainly in mesenchymal cells and in neural tissue.

摘要

从海鞘肠鳃纲幼虫制备的cDNA文库中分离出一种编码RNA结合蛋白的cDNA。推测的162个氨基酸的蛋白质在N端区域包含一个共有序列RNA结合结构域的拷贝,在C端区域包含一个富含甘氨酸的结构域。体外翻译的蛋白质能结合各种RNA同聚物,优先结合聚尿嘧啶、聚腺嘌呤和聚鸟嘌呤,并且结合受到离子强度增加的影响。Northern印迹分析显示有一个约0.7 kb长的单一转录本,在胚胎发育期间存在,在原肠胚和幼虫阶段有两个主要积累峰。对不同发育阶段胚胎进行的整体原位杂交实验表明,该基因主要在间充质细胞和神经组织中表达。

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