ANTIBODY-SELECTABLE FILAMENTOUS FD PHAGE VECTORS - AFFINITY PURIFICATION OF TARGET GENES
ANTIBODY-SELECTABLE FILAMENTOUS FD PHAGE VECTORS - AFFINITY PURIFICATION OF TARGET GENES
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DOI：
10.1016/0378-1119(88)90495-7
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发表时间：
1988-12-20
期刊：
影响因子：
3.5
通讯作者：
SMITH, GP
中科院分区：
文献类型：
--
作者：
PARMLEY, SF;SMITH, GP
外源DNA片段可以插入到丝状噬菌体的次要外壳蛋白基因中，产生融合蛋白，该融合蛋白被并入病毒体中;我们将这些颗粒称为“融合噬菌体”。外源氨基酸展示在表面上，允许携带来自靶基因的抗原决定簇的融合噬菌体通过对针对基因产物的抗体的亲和力以感染形式纯化。在这里，我们介绍融合噬菌体载体，接受外来的DNA插入与噬菌体功能的影响不大，并描述亲和纯化的病毒携带的目标决定簇从108倍过量的噬菌体不携带的决定簇，使用微量的抗体。这些“抗体选择性”载体是使用抗体克隆基因的常规表达系统的有希望的替代方案，尽管从实际文库中分离稀有克隆的能力仍有待证明。
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 acid 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 virins bearing a target determinant from a 108-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.