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Methods and Molecular Tools for Studying Endocytosis in Plants---an Overview

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Plant Endocytosis

Part of the book series: Plant Cell Monographs ((CELLMONO,volume 1))

Abstract

Proteins of the endocytosis machinery in plants, such as clathrin and adaptor proteins, were isolated and characterized using combinations of molecular biological (cloning and tagging) and biochemical methods (gel filtration, pull-down assays, surface plasmon resonance and immunoblotting). Other biochemical methods, such as cell fractionation and sucrose density gradients, were applied in order to isolate and further characterize clathrin-coated vesicles and endosomes in plants. Endocytosis was visualized in plant cells by using both non-fluorescent and fluorescent markers, and by employing antibodies raised against endosomal proteins or green fluorescent protein-tagged endocytic proteins in combination with diverse microscopic techniques, including confocal laser scanning microscopy and electron microscopy. Genetic and cell biological approaches were used together to address the role of a few proteins potentially involved in endocytosis. Additionally, biochemical and/or biophysical/electrophysiological methods were occasionally combined with microscopic methods (including both in situ and in vivo visualization) in plant endocytosis research.

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Acknowledgments

I thank Diedrik Menzel for critical reading of the manuscript and Ursulla Mettbach and Claudia Heym for excellent technical assistance, as well as to Mary Preuss and Erik Nielsen for providing Fig. 1a. This work was supported by a grant from the Slovak Grant Agency APVT (grant no. APVT-51-002302), Bratislava, Slovakia.

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  1. Institute of Cellular and Molecular Botany, University of Bonn, Kirschallee 1, 53115, Bonn, Germany

    Jozef Šamaj

  2. Institute of Plant Genetics and Biotechnology, Slovak Academy of Sciences, Akademicka 2, 949 01, Nitra, Slovakia

    Jozef Šamaj

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Correspondence to Jozef Šamaj .

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Jozef Šamaj František Baluška Diedrik Menzel

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Šamaj, J. Methods and Molecular Tools for Studying Endocytosis in Plants---an Overview. In: Šamaj, J., Baluška, F., Menzel, D. (eds) Plant Endocytosis. Plant Cell Monographs, vol 1. Springer, Berlin, Heidelberg. https://doi.org/10.1007/7089_002

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