编码大鼠脂皮质素I（钙结合蛋白II）的cDNA在大肠杆菌中的分子克隆及表达。

Molecular cloning and expression in Escherichia coli of the cDNA coding for rat lipocortin I (calpactin II).

作者信息

Shimizu Y, Takabayashi E, Yano S Y, Shimizu N, Yamada K, Gushima H

机构信息

Molecular Biology Department, Yamanouchi Pharmaceutical Co. Ltd., Tokyo, Japan.

出版信息

Gene. 1988 May 15;65(1):141-7. doi: 10.1016/0378-1119(88)90427-1.

DOI:10.1016/0378-1119(88)90427-1

PMID:2969352

Abstract

Lipocortins (LC) are a family of proteins that were initially described to be induced by glucocorticosteroids and to inhibit phospholipase A2 (PLA2). Using oligodeoxynucleotide probes corresponding to partial amino acid (aa) sequences of rat lipocortin I (LCI), we have isolated a cDNA clone for rat LCI from a cDNA library prepared from poly(A)+RNA of peritoneal cells of dexamethasone-treated rat. The cDNA insert (1355 bp) had an open reading frame of 1038 bp that encoded a 346-aa polypeptide (Mr 38,784). The nucleotide sequence and the amino acid sequence deduced from it showed high homology with the reported sequences of human LCI. A plasmid containing the trc promoter and cDNA sequence for 346 aa residues of the rat LCI was constructed and expressed in Escherichia coli. Antibody to human LCI crossreacted with the recombinant rat LCI, and the recombinant protein had characteristics of natural rat LCI including PLA2 inhibitory activity in vitro.

摘要

脂皮质素（LC）是一类蛋白质，最初被描述为由糖皮质激素诱导并抑制磷脂酶A2（PLA2）。利用与大鼠脂皮质素I（LCI）部分氨基酸（aa）序列相对应的寡脱氧核苷酸探针，我们从用 dexamethasone 处理的大鼠腹膜细胞的 poly(A)+RNA 制备的 cDNA 文库中分离出大鼠 LCI 的 cDNA 克隆。该 cDNA 插入片段（1355 bp）有一个 1038 bp 的开放阅读框，编码一个 346 个氨基酸的多肽（Mr 38,784）。其核苷酸序列及由此推导的氨基酸序列与已报道的人 LCI 序列具有高度同源性。构建了一个含有 trc 启动子和大鼠 LCI 346 个氨基酸残基 cDNA 序列的质粒，并在大肠杆菌中表达。抗人 LCI 抗体与重组大鼠 LCI 发生交叉反应，该重组蛋白具有天然大鼠 LCI 的特性，包括体外 PLA2 抑制活性。