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一种用于植物高通量基因功能分析的灵活克隆兼容载体系统。

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

机构信息

Key Laboratory of Biology and Genetic Resources of Tropical Crops, Sanya Research Institute, Institute of Tropical Biosciences and Biotechnology, Chinese Academy of Tropical Agricultural Science & Key Laboratory for Biology and Genetic Resources of Tropical Crops of Hainan Province, Hainan Institute of Tropical Agricultural Resources, Haikou 571101, China.

Key Laboratory of Biology and Genetic Resources of Tropical Crops, Sanya Research Institute, Institute of Tropical Biosciences and Biotechnology, Chinese Academy of Tropical Agricultural Science & Key Laboratory for Biology and Genetic Resources of Tropical Crops of Hainan Province, Hainan Institute of Tropical Agricultural Resources, Haikou 571101, China.

出版信息

Plant Commun. 2023 Mar 13;4(2):100471. doi: 10.1016/j.xplc.2022.100471. Epub 2022 Nov 9.

DOI:10.1016/j.xplc.2022.100471
PMID:36352791
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10030367/
Abstract

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.

摘要

植物表达载体是基因功能分析和分子植物育种的重要工具。通过分子克隆技术将目的基因转移到载体上。Nimble Cloning 是一种新开发的分子克隆方法，具有简单、高效和标准化的优点。本研究开发了一个包含 55 个 Nimble Cloning 兼容载体的“pNC”载体系统，用于植物基因的功能分析。这些载体包含由独特接头侧翼的 NC 框，用于一步法和标准化的 Nimble Cloning。我们证明，pNC 载体对于植物基因的功能分析非常方便和有效，包括基因异位表达、蛋白质亚细胞定位、蛋白质-蛋白质相互作用、基因沉默（RNAi）、病毒诱导的基因沉默、启动子活性和 CRISPR-Cas9 介导的基因组编辑的研究。“pNC”载体系统代表了一种高通量工具包，可以促进植物功能基因组学的大规模分析。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c2b5/10030367/7b7dfad47053/gr7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c2b5/10030367/852b8c7afa6c/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c2b5/10030367/4982b528b398/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c2b5/10030367/b6579e970297/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c2b5/10030367/3d0fe15a5e8c/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c2b5/10030367/0cb8e163a424/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c2b5/10030367/361ab229afed/gr6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c2b5/10030367/7b7dfad47053/gr7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c2b5/10030367/852b8c7afa6c/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c2b5/10030367/4982b528b398/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c2b5/10030367/b6579e970297/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c2b5/10030367/3d0fe15a5e8c/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c2b5/10030367/0cb8e163a424/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c2b5/10030367/361ab229afed/gr6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c2b5/10030367/7b7dfad47053/gr7.jpg

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