内皮素转化酶活性向CHO-K1细胞的基因转移：通过反向溶血空斑试验检测阳性细胞。

Genetic transfer of endothelin converting enzyme activity to CHO-K1 cells: detection of positive cells by reverse hemolytic plaque assay.

作者信息

Shiraki T, Sawamura T, Ikura T, Kobayashi S, Miwa S, Masaki T

机构信息

Department of Pharmacology, Faculty of Medicine, Kyoto University, Japan.

出版信息

FEBS Lett. 1994 Sep 5;351(2):197-200. doi: 10.1016/0014-5793(94)00830-2.

DOI:10.1016/0014-5793(94)00830-2

PMID:8082764

Abstract

We have established a novel method of molecular cloning of endothelin converting enzyme, a key enzyme in the production of a potent vasoconstrictor endothelin-1, by modification of the reverse hemolytic plaque assay. Also, we demonstrated that a cell line, CHO-K1, showed no detectable activity of endothelin converting enzyme. This cell line was transfected with a cDNA library of bovine endothelial cells. The modified reverse hemolytic plaque assay was shown to detect even a single CHO-K1 cell that was changed to produce mature ET-1 by transfection. Thus, this novel method is suggested to be useful for the molecular cloning of other secreted antigens and their processing enzyme.

摘要

我们通过改进反向溶血空斑试验，建立了一种新的分子克隆内皮素转化酶的方法，内皮素转化酶是强效血管收缩剂内皮素-1生成过程中的关键酶。此外，我们还证明了一种细胞系CHO-K1未显示出可检测到的内皮素转化酶活性。用牛内皮细胞的cDNA文库转染该细胞系。结果表明，改进后的反向溶血空斑试验甚至可以检测到单个通过转染而转变为能产生成熟ET-1的CHO-K1细胞。因此，这种新方法被认为可用于其他分泌性抗原及其加工酶的分子克隆。