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人载脂蛋白A-II:前体脱辅基脂蛋白A-II的完整核酸序列。

Human apolipoprotein A-II: complete nucleic acid sequence of preproapo A-II.

作者信息

Lackner K J, Law S W, Brewer H B

出版信息

FEBS Lett. 1984 Sep 17;175(1):159-64. doi: 10.1016/0014-5793(84)80590-6.

Abstract

The complete cDNA nucleic acid sequence of preproapolipoprotein (apo) A-II, a major protein constituent of high density lipoproteins, has been determined on clones from a human liver ds-cDNA library. Clones containing ds-cDNA for apoA-II were identified in the human liver ds-cDNA library using synthetic oligonucleotides as probes. Of 3200 clones screened, 4 reacted with the oligonucleotide probes. The DNA sequence coding for amino acids -17 to +17 of apoA-II were determined by Maxam-Gilbert sequence analysis of restriction fragments isolated from one of these clones, pMDB2049. The remainder of the cDNA sequence was established by sequence analysis of a primer extension product synthesized utilizing a restriction fragment near the 5'-end of clone pMDB2049 as primer with total liver mRNA. The apoA-II mRNA encodes for a 100 amino acid protein, preproapoA-II that has an 18 amino acid prepeptide and a 5 amino acid propeptide terminating with a basic dipeptide (Arg-Arg) at the cleavage site to mature apoA-II.

摘要

已在来自人肝脏双链cDNA文库的克隆上确定了前原载脂蛋白(apo)A-II的完整cDNA核酸序列,apo A-II是高密度脂蛋白的主要蛋白质成分。使用合成寡核苷酸作为探针,在人肝脏双链cDNA文库中鉴定出含有apo A-II双链cDNA的克隆。在筛选的3200个克隆中,有4个与寡核苷酸探针发生反应。通过对从其中一个克隆pMDB2049分离的限制性片段进行Maxam-Gilbert序列分析,确定了编码apo A-II氨基酸-17至+17的DNA序列。利用克隆pMDB2049 5'-末端附近的限制性片段作为引物与总肝脏mRNA合成的引物延伸产物进行序列分析,确定了cDNA序列的其余部分。apo A-II mRNA编码一种100个氨基酸的蛋白质,即前原apo A-II,它有一个18个氨基酸的前肽和一个5个氨基酸的前体肽,在成熟apo A-II的切割位点以碱性二肽(Arg-Arg)终止。

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