A Nimble Cloning-compatible vector system for high-throughput gene functional analysis in plants

Plant expression vectors are essential tools for gene functional analysis and molecular plant breeding. The gene of interest is transferred to the vector by molecular cloning technology. Nimble Cloning is a newly developed molecular cloning method with the advantages of simplicity, efficiency, and s...

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Autores principales: Yan, Pu, Tuo, Decai, Shen, Wentao, Deng, Haida, Zhou, Peng, Gao, Xinzheng
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10030367/
https://www.ncbi.nlm.nih.gov/pubmed/36352791
http://dx.doi.org/10.1016/j.xplc.2022.100471
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author Yan, Pu
Tuo, Decai
Shen, Wentao
Deng, Haida
Zhou, Peng
Gao, Xinzheng
author_facet Yan, Pu
Tuo, Decai
Shen, Wentao
Deng, Haida
Zhou, Peng
Gao, Xinzheng
author_sort Yan, Pu
collection PubMed
description Plant expression vectors are essential tools for gene functional analysis and molecular plant breeding. The gene of interest is transferred to the vector by molecular cloning technology. Nimble Cloning is a newly developed molecular cloning method with the advantages of simplicity, efficiency, and standardization. In this study, we developed a “pNC” vector system that contains 55 Nimble Cloning-compatible vectors for functional analysis of genes in plants. These vectors contain the NC frame flanked by unique adapters for one-step and standardized Nimble Cloning. We demonstrate that the pNC vectors are convenient and effective for the functional analysis of plant genes, including the study of gene ectopic expression, protein subcellular localization, protein–protein interaction, gene silencing (RNAi), virus-induced gene silencing, promoter activity, and CRISPR–Cas9-mediated genome editing. The “pNC” vector system represents a high-throughput toolkit that can facilitate the large-scale analysis of plant functional genomics.
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spelling pubmed-100303672023-03-23 A Nimble Cloning-compatible vector system for high-throughput gene functional analysis in plants Yan, Pu Tuo, Decai Shen, Wentao Deng, Haida Zhou, Peng Gao, Xinzheng Plant Commun Resource Article Plant expression vectors are essential tools for gene functional analysis and molecular plant breeding. The gene of interest is transferred to the vector by molecular cloning technology. Nimble Cloning is a newly developed molecular cloning method with the advantages of simplicity, efficiency, and standardization. In this study, we developed a “pNC” vector system that contains 55 Nimble Cloning-compatible vectors for functional analysis of genes in plants. These vectors contain the NC frame flanked by unique adapters for one-step and standardized Nimble Cloning. We demonstrate that the pNC vectors are convenient and effective for the functional analysis of plant genes, including the study of gene ectopic expression, protein subcellular localization, protein–protein interaction, gene silencing (RNAi), virus-induced gene silencing, promoter activity, and CRISPR–Cas9-mediated genome editing. The “pNC” vector system represents a high-throughput toolkit that can facilitate the large-scale analysis of plant functional genomics. Elsevier 2022-11-09 /pmc/articles/PMC10030367/ /pubmed/36352791 http://dx.doi.org/10.1016/j.xplc.2022.100471 Text en © 2022 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Resource Article
Yan, Pu
Tuo, Decai
Shen, Wentao
Deng, Haida
Zhou, Peng
Gao, Xinzheng
A Nimble Cloning-compatible vector system for high-throughput gene functional analysis in plants
title A Nimble Cloning-compatible vector system for high-throughput gene functional analysis in plants
title_full A Nimble Cloning-compatible vector system for high-throughput gene functional analysis in plants
title_fullStr A Nimble Cloning-compatible vector system for high-throughput gene functional analysis in plants
title_full_unstemmed A Nimble Cloning-compatible vector system for high-throughput gene functional analysis in plants
title_short A Nimble Cloning-compatible vector system for high-throughput gene functional analysis in plants
title_sort nimble cloning-compatible vector system for high-throughput gene functional analysis in plants
topic Resource Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10030367/
https://www.ncbi.nlm.nih.gov/pubmed/36352791
http://dx.doi.org/10.1016/j.xplc.2022.100471
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