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Wiley InterScience | |||||||||
  The Plant JournalVolume 52 Issue 1, Pages 185 - 195 Published Online: 30 Jul 2007 Journal compilation © 2010 Blackwell Publishing Ltd and the Society for Experimental Biology Published in association with the Society for Experimental Biology
Abstract | References | Full Text: HTML, PDF (Size: 336K) | Supporting Information | Related Articles | Citation Tracking  TECHNICAL ADVANCE Split luciferase complementation assay to study protein–protein interactions in Arabidopsis protoplasts Copyright 2007 The Authors Journal compilation 2007 Blackwell Publishing Ltd KEYWORDS protein–protein interactions • split luciferase •
in vivo
• protoplasts •
Arabidopsis
• FlAsH ABSTRACTWe 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. Received 21 February 2007; revised 29 May 2007; accepted 5 June 2007. | 
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