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绵羊肾素的序列及组织表达

The sequence and tissue expression of ovine renin.

作者信息

Aldred G P, Fu P, Crawford R J, Fernley R T

机构信息

Howard Florey Institute of Experimental Physiology and Medicine, Parkville, Victoria, Australia.

出版信息

J Mol Endocrinol. 1992 Feb;8(1):3-11. doi: 10.1677/jme.0.0080003.

Abstract

The primary structure of the sheep renin precursor has been determined from its cDNA sequence. A library of cDNA clones was constructed from adrenalectomized sheep kidney poly(A)+ RNA and screened for sheep renin sequences with a cloned mouse renin cDNA probe. Of the 300,000 clones generated, 24 were hybridization positive and the nucleotide sequences of two of the longest clones were determined. These clones coded for the mature sheep renin protein and the 3'-untranslated sequence but did not extend to the amino-terminal region of preprorenin. Clones corresponding to the 5' region of renin mRNA were generated by the polymerase chain reaction and their nucleotide sequences determined. The sheep renin precursor consists of 400 amino acids with a putative leader sequence of 14 amino acids and a putative 45 or 53 amino acid prosegment. The mature sheep renin protein has a 73% sequence identity with human renin. Northern analysis demonstrated the presence of renin mRNA in the kidney but not in other tissues in the sheep. While sodium depletion of sheep caused a rise in renin mRNA in the kidney, adrenalectomy also led to a large increase in renal renin mRNA. Southern analysis of genomic DNA suggests that there is only one gene coding for renin in the sheep.

摘要

绵羊肾素前体的一级结构已通过其cDNA序列得以确定。从肾上腺切除的绵羊肾脏的多聚腺苷酸加尾RNA构建了一个cDNA克隆文库,并用克隆的小鼠肾素cDNA探针筛选绵羊肾素序列。在产生的300,000个克隆中,有24个杂交呈阳性,并测定了两个最长克隆的核苷酸序列。这些克隆编码成熟的绵羊肾素蛋白和3'非翻译序列,但未延伸至前肾素原的氨基末端区域。通过聚合酶链反应产生了与肾素mRNA 5'区域相对应的克隆,并测定了它们的核苷酸序列。绵羊肾素前体由400个氨基酸组成,具有一个推定的14个氨基酸的前导序列和一个推定的45或53个氨基酸的前肽段。成熟的绵羊肾素蛋白与人类肾素的序列同一性为73%。Northern分析表明,绵羊肾脏中存在肾素mRNA,而其他组织中则没有。虽然绵羊的钠缺乏导致肾脏中肾素mRNA升高,但肾上腺切除也导致肾脏肾素mRNA大幅增加。基因组DNA的Southern分析表明,绵羊中只有一个编码肾素的基因。

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