期刊
ANALYTICAL BIOCHEMISTRY
卷 434, 期 1, 页码 93-95出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ab.2012.10.041
关键词
Pull-down assay; Protein-protein interaction; In vitro selection; Puromycin; Cell-free translation
资金
- JSPS KAKENHI [23510253]
- Grants-in-Aid for Scientific Research [23510253, 23659141] Funding Source: KAKEN
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.
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