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人肾素前体cDNA的克隆与序列分析

Cloning and sequence analysis of cDNA for human renin precursor.

作者信息

Imai T, Miyazaki H, Hirose S, Hori H, Hayashi T, Kageyama R, Ohkubo H, Nakanishi S, Murakami K

出版信息

Proc Natl Acad Sci U S A. 1983 Dec;80(24):7405-9. doi: 10.1073/pnas.80.24.7405.

Abstract

The primary structure of human renin precursor has been deduced from its cDNA sequence. A library of cDNA clones was constructed from human kidney poly(A)+ RNA by applying the vector/primer method of Okayama and Berg. The library was screened for human renin sequences by hybridization with the previously cloned mouse renin cDNA. Of the 240,000 colonies screened, 35 colonies that were positive for hybridization were isolated. Two recombinant plasmids containing long inserts of about 1,300 and 1,600 base pairs were selected for sequence analysis. The amino acid sequence predicted from the cDNA sequence shows that the human renin precursor consists of 406 amino acids with a pre and a pro segment carrying 20 and 46 amino acids, respectively. A high degree of sequence homology was found upon comparison of the mouse and human renins. Close similarities were also observed in the primary structures of renin and aspartyl proteinases that have known three-dimensional structures, suggesting a similar tertiary structure for renin.

摘要

人肾素前体的一级结构已根据其cDNA序列推导得出。采用冈山县和伯格的载体/引物法,从人肾多聚腺苷酸加尾RNA构建了一个cDNA克隆文库。通过与先前克隆的小鼠肾素cDNA杂交,筛选该文库中的人肾素序列。在筛选的240,000个菌落中,分离出35个杂交阳性菌落。选择了两个含有约1300和1600个碱基对长插入片段的重组质粒进行序列分析。从cDNA序列预测的氨基酸序列表明,人肾素前体由406个氨基酸组成,其前肽和前原肽分别含有20个和46个氨基酸。比较小鼠和人肾素时发现了高度的序列同源性。在已知三维结构的肾素和天冬氨酸蛋白酶的一级结构中也观察到了密切的相似性,这表明肾素具有相似的三级结构。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fc9c/389959/8a481165900d/pnas00650-0041-a.jpg

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