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Tissue-specific isolation of tagged Arabidopsis plastids
Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Institutionen för fysiologisk botanik. Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Umeå Plant Science Centre (UPSC).ORCID-id: 0000-0001-8313-3535
Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Institutionen för fysiologisk botanik. Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Umeå Plant Science Centre (UPSC).ORCID-id: 0000-0002-0546-7721
2023 (engelsk)Inngår i: Current protocols, E-ISSN 2691-1299, Vol. 3, nr 2, artikkel-id e673Artikkel i tidsskrift (Fagfellevurdert) Published
Abstract [en]

Plastids are found in all plant cell types. However, most extraction methods to study these organelles are performed at the organ level (e.g., leaf, root, fruit) and do not allow for tissue-specific resolution, which hinders our understanding of their physiology. Therefore, IPTACT (Isolation of Plastids TAgged in specific Cell Types) was developed to isolate plastids in a tissue-specific manner in Arabidopsis thaliana (Arabidopsis). Plastids are biotinylated using one-shot transgenic lines, and tissue specificity is achieved with a suitable promoter as long as such a promoter exists. Cell-specific biotinylated plastids are then isolated with 2.8-µm streptavidin beads. Plastids extracted by IPTACT are suitable for RNA or protein isolation and subsequent tissue-specific OMICs analyses. This method provides the user with a powerful tool to investigate plastidial functions at cell-type resolution. Furthermore, it can easily be combined with studies using diverse genetic backgrounds and/or different developmental or stress conditions.

sted, utgiver, år, opplag, sider
John Wiley & Sons, 2023. Vol. 3, nr 2, artikkel-id e673
Emneord [en]
Arabidopsis, biotin-streptavidin interaction, editable Golden Gate plasmids, plastids, tissue-specific isolation
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Identifikatorer
URN: urn:nbn:se:umu:diva-205383DOI: 10.1002/cpz1.673ISI: 001282191900027PubMedID: 36799650Scopus ID: 2-s2.0-85148334462OAI: oai:DiVA.org:umu-205383DiVA, id: diva2:1745464
Forskningsfinansiär
The Royal Swedish Academy of Sciences, BS2021‐0001Swedish Foundation for Strategic Research, FFF20‐0008VinnovaKnut and Alice Wallenberg FoundationTilgjengelig fra: 2023-03-23 Laget: 2023-03-23 Sist oppdatert: 2025-04-24bibliografisk kontrollert

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