A pull-down method with a biotinylated bait protein prepared by cell-free translation using a puromycin linker

被引：10

作者：

Mochizuki, Yuki ^{[1
]}

Kohno, Fumiaki ^{[1
]}

Nishigaki, Koichi ^{[1
]}

Nemoto, Naoto ^{[1
]}

机构：

[1] Saitama Univ, Grad Sch Sci & Engn, Sakura Ku, Saitama 3388570, Japan

来源：

ANALYTICAL BIOCHEMISTRY | 2013年 / 434卷 / 01期

关键词：

Pull-down assay; Protein-protein interaction; In vitro selection; Puromycin; Cell-free translation; EPITOPE; ASSAYS;

D O I：

10.1016/j.ab.2012.10.041

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

In this paper, we demonstrate a novel pull-down method that dramatically reduces the cost and preparation time of a bait protein by cell-free translation with a puromycin linker. With the C-terminus of the bait protein linked to biotin through a puromycin molecule after the translation reaction and subsequent mRNA degradation by RNase, the prey protein was easily pulled down by streptavidin-coated magnetic beads in a test tube. Three fluorescent prey protein types were tested and confirmed by gel electrophoresis to be pulled down easily and rapidly, depending on their affinity. (C) 2012 Elsevier Inc. All rights reserved.

引用

页码：93 / 95

页数：3

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