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Wageningen University

5 ARTICLES PUBLISHED IN JoVE

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Immunology and Infection

Avian Influenza Surveillance with FTA Cards: Field Methods, Biosafety, and Transportation Issues Solved
Robert H.S. Kraus 1, Pim van Hooft 1, Jonas Waldenström 2, Neus Latorre-Margalef 2, Ronald C. Ydenberg 1,3, Herbert H.T. Prins 1
1Resource Ecology Group, Wageningen University, 2Section for Zoonotic Ecology and Epidemiology, School of Natural Sciences, Linnaeus University, 3Centre for Wildlife Ecology, Simon Fraser University

A method to preserve, detect and sequence RNA from Avian Influenza Viruses was validated and extended using natural faecal samples from birds. This technique removes the necessity of maintaining a cool chain and handling of infectious viruses and can be applied in a 96-well high-throughput setup.

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Biology

Agroinfiltration and PVX Agroinfection in Potato and Nicotiana benthamiana
Juan Du 1,2, Hendrik Rietman 1, Vivianne G. A. A. Vleeshouwers 1
1Wageningen UR Plant Breeding, Wageningen University, 2Key Laboratory of Horticultural Plant Biology, Huazhong Agricultural University, Ministry of Education, National Centre for Vegetable Improvement (Central China), Potato Engineering and Technology Research Centre of Hubei Province, Huazhong Agricultural University

Agroinfiltration and PVX agroinfection are routine functional assays for transient ectopic expression of genes in plants. These methods are efficient assays in effectoromics strategies (rapid resistance and avirulence gene discovery) and crucial to modern research in molecular plant pathology. They meet the demand for robust high-throughput functional analysis in plants.

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Environment

Modification and Application of a Leaf Blower-vac for Field Sampling of Arthropods
Yi Zou 1, Mario D. van Telgen 1, Junhui Chen 2, Haijun Xiao 2, Joop de Kraker 3, Felix J. J. A. Bianchi 4, Wopke van der Werf 1
1Centre for Crop Systems Analysis, Wageningen University, 2Institute of Entomology, Jiangxi Agricultural University, 3Department of Science, Open University, 4Farming Systems Ecology, Wageningen University

Assessment of arthropod abundance in crops is critical for investigating population dynamics and species interactions. Here we describe the modification and application of a leaf blower-vac for suction sampling of arthropods in rice.

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Cancer Research

Detection of Mitochondria Membrane Potential to Study CLIC4 Knockdown-induced HN4 Cell Apoptosis In Vitro
Jinsen Lu 1, Lele Wu 2, Xiaoke Wang 2, Jinhang Zhu 2, Juan Du 2, Bing Shen 2
1School of Basic Medical Sciences, Anhui Medical University, 2Department of Orthopedics, Anhui Provincial Hospital, Anhui Medical University

Here we present a detailed protocol for the application of rhodamine 123 to identify the mitochondrial membrane potential (MMP) and study CLIC4 knockdown-induced HN4 cell apoptosis in vitro. Under common fluorescence microscope and confocal laser scanning fluorescence microscope, the real-time change of the MMP was recorded.

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Biochemistry

Expression and Purification of the Human Lipid-sensitive Cation Channel TRPC3 for Structural Determination by Single-particle Cryo-electron Microscopy
Emery Haley *1, Wooyoung Choi *1, Chen Fan 1, Weinan Sun 2,3, Juan Du 1, Wei Lü 1
1Van Andel Institute, 2Vollum Institute, 3Janelia Research Campus

This protocol describes techniques used to determine ion channel structures by cryo-electron microscopy, including a baculovirus system used to efficiently express genes in mammalian cells with minimum effort and toxicity, protein extraction, purification, and quality checking, sample grid preparation and screening, as well as data collection and processing.

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