An improved toolkit of gateway- and gibson assembly-compatible vectors for protoplast transfection and agrobacterium-mediated plant transformation

Abstract Objective Understanding the regulation and function of plant genes is essential for addressing the challenges faced by modern agriculture. Plant transformation, in conjunction with fluorescence microscopy, offers a powerful approach to investigate the dynamic behavior of plant genes and the...

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Main Authors: Jeffrey R. Allen, Michael Flanagan, Sunita Pathak, Samantha K. Emenecker, Ryan J. Emenecker, Lucia C. Strader
Format: Article
Language:English
Published: BMC 2025-04-01
Series:BMC Research Notes
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Online Access:https://doi.org/10.1186/s13104-025-07231-1
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author Jeffrey R. Allen
Michael Flanagan
Sunita Pathak
Samantha K. Emenecker
Ryan J. Emenecker
Lucia C. Strader
author_facet Jeffrey R. Allen
Michael Flanagan
Sunita Pathak
Samantha K. Emenecker
Ryan J. Emenecker
Lucia C. Strader
author_sort Jeffrey R. Allen
collection DOAJ
description Abstract Objective Understanding the regulation and function of plant genes is essential for addressing the challenges faced by modern agriculture. Plant transformation, in conjunction with fluorescence microscopy, offers a powerful approach to investigate the dynamic behavior of plant genes and the proteins they encode. We previously developed a set of Gateway-compatible tissue-specific plant transformation vectors. In this paper we aim to expand the toolkit of vectors available for Agrobacterium-mediated plant transformation and protoplast transfection. Results Here, we introduce new Agrobacterium-mediated plant transformation vectors by introducing additional fluorophores to create the pJRA vector series. Additionally, we introduce the pLCS series of vectors, a new set of modular Gateway- and Gibson assembly-compatible vectors designed for protoplast transfection. All described vectors are available from Addgene to serve as a resource for the plant research community.
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spelling doaj-art-fee3482d83fb4877a82ba9c1d344eb7b2025-08-20T02:11:58ZengBMCBMC Research Notes1756-05002025-04-011811810.1186/s13104-025-07231-1An improved toolkit of gateway- and gibson assembly-compatible vectors for protoplast transfection and agrobacterium-mediated plant transformationJeffrey R. Allen0Michael Flanagan1Sunita Pathak2Samantha K. Emenecker3Ryan J. Emenecker4Lucia C. Strader5Department of Biology, Duke UniversityDepartment of Biology, Duke UniversityDepartment of Biology, Duke UniversityDepartment of Biology, Washington University in St. LouisDepartment of Biology, Washington University in St. LouisDepartment of Biology, Duke UniversityAbstract Objective Understanding the regulation and function of plant genes is essential for addressing the challenges faced by modern agriculture. Plant transformation, in conjunction with fluorescence microscopy, offers a powerful approach to investigate the dynamic behavior of plant genes and the proteins they encode. We previously developed a set of Gateway-compatible tissue-specific plant transformation vectors. In this paper we aim to expand the toolkit of vectors available for Agrobacterium-mediated plant transformation and protoplast transfection. Results Here, we introduce new Agrobacterium-mediated plant transformation vectors by introducing additional fluorophores to create the pJRA vector series. Additionally, we introduce the pLCS series of vectors, a new set of modular Gateway- and Gibson assembly-compatible vectors designed for protoplast transfection. All described vectors are available from Addgene to serve as a resource for the plant research community.https://doi.org/10.1186/s13104-025-07231-1Protoplast transfectionAgrobacterium transformationTransgenic plantPlant transformationTransient expressionLive cell imaging
spellingShingle Jeffrey R. Allen
Michael Flanagan
Sunita Pathak
Samantha K. Emenecker
Ryan J. Emenecker
Lucia C. Strader
An improved toolkit of gateway- and gibson assembly-compatible vectors for protoplast transfection and agrobacterium-mediated plant transformation
BMC Research Notes
Protoplast transfection
Agrobacterium transformation
Transgenic plant
Plant transformation
Transient expression
Live cell imaging
title An improved toolkit of gateway- and gibson assembly-compatible vectors for protoplast transfection and agrobacterium-mediated plant transformation
title_full An improved toolkit of gateway- and gibson assembly-compatible vectors for protoplast transfection and agrobacterium-mediated plant transformation
title_fullStr An improved toolkit of gateway- and gibson assembly-compatible vectors for protoplast transfection and agrobacterium-mediated plant transformation
title_full_unstemmed An improved toolkit of gateway- and gibson assembly-compatible vectors for protoplast transfection and agrobacterium-mediated plant transformation
title_short An improved toolkit of gateway- and gibson assembly-compatible vectors for protoplast transfection and agrobacterium-mediated plant transformation
title_sort improved toolkit of gateway and gibson assembly compatible vectors for protoplast transfection and agrobacterium mediated plant transformation
topic Protoplast transfection
Agrobacterium transformation
Transgenic plant
Plant transformation
Transient expression
Live cell imaging
url https://doi.org/10.1186/s13104-025-07231-1
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