Person:
Solís González, María Teresa

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First Name
María Teresa
Last Name
Solís González
URI
https://hdl.handle.net/20.500.14352/74327
Affiliation
Universidad Complutense de Madrid
Faculty / Institute
Ciencias Biológicas
Department
Genética, Fisiología y Microbiología
Area
Fisiología Vegetal
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Author: Sánchez Testillano, Pilar × Subject: 577.112 ×

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    In situ/subcellular localization of arabinogalactan protein expression by fluorescent in situ hybridization (FISH)
    (The plant cell wall, 2020) Da Costa, Mário Luís; Solís González, María Teresa; Sánchez Testillano, Pilar; Coimbra, Sílvia; Popper, Zoë A.
    The arabinogalactan proteins are highly glycosylated and ubiquitous in plants. They are involved in several aspects of plant development and reproduction; however, the mechanics behind their function remains for the most part unclear, as the carbohydrate moiety, covering the most part of the protein core, is poorly characterized at the individual protein level. Traditional immunolocalization using antibodies that recognize the glycosidic moiety of the protein cannot be used to elucidate individual proteins’ distribution, function, or interactors. Indirect approaches are typically used to study these proteins, relying on reverse genetic analysis of null mutants or using a reporter fusion system. In the method presented here, we propose the use of RNA probes to assist in the localization of individual AGPs expression/mRNAs in tissues of Arabidopsis by fluorescent in situ hybridization, FISH. An extensive description of all aspects of this technique is provided, from RNA probe synthesis to the hybridization, trying to overcome the lack of specific antibodies for the protein core of AGPs.