3 articles published in JoVE
Real-time In Vivo Recording of Arabidopsis Calcium Signals During Insect Feeding Using a Fluorescent Biosensor Thomas R. Vincent1, James Canham1, Masatsugu Toyota2,3,4, Marieta Avramova1, Sam T. Mugford5, Simon Gilroy2, Anthony J. Miller1, Saskia Hogenhout5, Dale Sanders1 1Department of Metabolic Biology, John Innes Centre, Norwich Research Park, 2Department of Botany, University of Wisconsin, Madison, 3Department of Biochemistry and Molecular Biology, Saitama University, 4Precursory Research for Embryonic Science and Technology (PRESTO), Japan Science and Technology Agency (JST), 5Department of Cell and Developmental Biology, John Innes Centre, Norwich Research Park This protocol outlines a simple method for analyzing calcium signals in plants generated by feeding hemipteran insects, such as aphids. Arabidopsis thaliana transformed with the GFP calcium biosensor GCaMP3 allow for the real-time in vivo imaging of calcium dynamics with a high temporal and spatial resolution.
Purification of High Molecular Weight Genomic DNA from Powdery Mildew for Long-Read Sequencing Joanna M. Feehan*1,2, Katherine E. Scheibel*1, Salim Bourras3, William Underwood4, Beat Keller3, Shauna C. Somerville1 1Department of Plant and Microbial Biology, University of California Berkeley, 2John Innes Centre, Norwich Research Park, 3Department of Plant and Microbial Biology, University of Zürich, 4USDA-ARS Sunflower and Plant Biology Research Unit Described here is a method for the extraction, purification, and quality control of genomic DNA from the obligate biotrophic fungal pathogen, powdery mildew, for use in long-read genome sequencing.
Identification of Post-translational Modifications of Plant Protein Complexes Sophie J. M. Piquerez1, Alexi L. Balmuth2, Jan Sklenář2, Alexandra M.E. Jones1,2, John P. Rathjen3, Vardis Ntoukakis1 1School of Life Sciences, University of Warwick, 2The Sainsbury Laboratory, Norwich Research Park, 3Research School of Biology, The Australian National University We describe here a protocol for the purification and characterization of plant protein complexes. We demonstrate that by immunoprecipitating a single protein within a complex, so we can identify its post-translational modifications and its interacting partners.