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通过高效COS细胞表达系统对CD28互补DNA进行分子克隆。

Molecular cloning of a CD28 cDNA by a high-efficiency COS cell expression system.

作者信息

Aruffo A, Seed B

机构信息

Department of Molecular Biology, Massachusetts General Hospital, Boston 02114.

出版信息

Proc Natl Acad Sci U S A. 1987 Dec;84(23):8573-7. doi: 10.1073/pnas.84.23.8573.

Abstract

CD28 (Tp44) is a human T-cell-specific homodimer surface protein that may participate in T-cell activation. We have isolated a cDNA clone encoding CD28 by a simple and highly efficient cloning strategy based on transient expression in COS cells. Central to this strategy is the use of an efficient method to prepare large plasmid cDNA libraries. The libraries are introduced into COS cells, where transient expression of surface antigen allows the isolation of cDNAs by way of monoclonal antibody binding. The CD28 cDNA encodes a highly glycosylated membrane protein with homology to the immunoglobulin superfamily and directs the production of a homodimer in transfected COS cells.

摘要

CD28(Tp44)是一种人类T细胞特异性同型二聚体表面蛋白,可能参与T细胞激活。我们通过基于COS细胞瞬时表达的简单高效克隆策略分离出了编码CD28的cDNA克隆。该策略的核心是使用一种高效方法制备大型质粒cDNA文库。将文库导入COS细胞,表面抗原的瞬时表达使得通过单克隆抗体结合分离cDNA成为可能。CD28 cDNA编码一种与免疫球蛋白超家族具有同源性的高度糖基化膜蛋白,并在转染的COS细胞中指导同型二聚体的产生。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cb5c/299587/2cff16981f5b/pnas00338-0410-a.jpg

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