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抗体可选择的丝状fd噬菌体载体:靶基因的亲和纯化

Antibody-selectable filamentous fd phage vectors: affinity purification of target genes.

作者信息

Parmley S F, Smith G P

机构信息

Division of Biological Sciences, University of Missouri, Columbia 65211.

出版信息

Gene. 1988 Dec 20;73(2):305-18. doi: 10.1016/0378-1119(88)90495-7.

Abstract

Foreign DNA fragments can be inserted into a minor coat protein gene of filamentous phage, creating a fusion protein that is incorporated into the virion; we call these particles "fusion phage". The foreign amino acids are displayed on the surface, allowing fusion phage bearing antigenic determinants from a target gene to be purified in infectious form by affinity to antibody directed against the gene product. Here we introduce fusion-phage vectors that accept foreign DNA inserts with little effect on phage function; and describe affinity purification of virions bearing a target determinant from a 10(8)-fold excess of phage not bearing the determinant, using minute amounts of antibody. These "antibody-selectable" vectors are a promising alternative to conventional expression systems for using antibodies to clone genes, though the ability to isolate rare clones from actual libraries remains to be demonstrated.

摘要

外源DNA片段可插入丝状噬菌体的次要外壳蛋白基因中,产生一种融合蛋白,该融合蛋白会被整合到病毒粒子中;我们将这些颗粒称为“融合噬菌体”。外源氨基酸展示在表面,使得携带来自靶基因的抗原决定簇的融合噬菌体能够通过与针对该基因产物的抗体的亲和力,以感染性形式被纯化。在此,我们介绍了融合噬菌体载体,其接受外源DNA插入片段时对噬菌体功能影响很小;并描述了使用微量抗体,从比携带靶决定簇的噬菌体多10⁸倍的不携带该决定簇的噬菌体中亲和纯化携带靶决定簇的病毒粒子。这些“抗体可选择”载体是用于利用抗体克隆基因的传统表达系统的一种有前景的替代方案,不过从实际文库中分离稀有克隆的能力仍有待证明。

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