Split luciferase complementation assay to study protein-protein interactions in Arabidopsis protoplasts

被引：109

作者：

Fujikawa, Yukichi ^{[1
]}

Kato, Naohiro ^{[1
]}

机构：

[1] Louisiana State Univ, Dept Biol Sci, Baton Rouge, LA 70803 USA

来源：

PLANT JOURNAL | 2007年 / 52卷 / 01期

关键词：

protein-protein interactions; split luciferase; in vivo; protoplasts; Arabidopsis; FlAsH;

D O I：

10.1111/j.1365-313X.2007.03214.x

中图分类号：

Q94 [植物学];

学科分类号：

071001 ;

摘要：

We developed a split luciferase complementation assay to study protein-protein interactions in Arabidopsis protoplasts. In this assay, the N- and C-terminal fragments of Renilla reniforms luciferase are translationally fused to bait and prey proteins, respectively. When the proteins interact, split luciferase becomes activated and emits luminescence that can be measured by a microplate luminometer. Split luciferase activity was measured by first transforming protoplasts with a DNA vector in a 96-well plate. DNA vector expressing both bait and prey genes was constructed through two independent in vitro DNA recombinant reactions, Gateway and Cre-loxP. As proof of concept, we detected the protein-protein interactions between the nuclear histones 2A and 2B, as well as between membrane proteins SYP (syntaxin of plant) 51 and SYP61, in Arabidopsis protoplasts.

引用

页码：185 / 195

页数：11

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