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Plant Biotechnology Journal
Article . 2025 . Peer-reviewed
License: CC BY NC ND
Data sources: Crossref
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Seamless, modular binary vectors for plant cell and molecular biology

Authors: Rory Greenhalgh; Wilson Horner; Katherine A. Klimpel; Snigdha Chatterjee; M. Regina Scarpin; Jacob O. Brunkard;

Seamless, modular binary vectors for plant cell and molecular biology

Abstract

SummaryIn plants, genetic manipulations are commonly performed using Agrobacterium tumefaciens, a plant pathogen whose abilities as a ‘natural genetic engineer’ have been co‐opted for biotechnological applications. In the widely used binary vector systems, Agrobacterium already contains a plasmid with most of the genes needed for virulence and is transformed with a second vector carrying the DNA of interest that will be transferred to the plant. Many widely used binary vectors have not been substantially improved during the past few decades, however, and often have legacy features that are not needed for many contemporary applications, such as cumbersome cloning sites that introduce sequence ‘scars’ between proteins of interest and any fusion protein, including GFP. Here, we introduce the assembly vector (AVEC) plasmids, a series of modular vector backbones that are easily customized using inexpensive DNA assembly cloning methods and that are ideal for testing multiple epitope tags, fluorophores, or non‐coding sequences, such as promoters or 5′/3′ untranslated regions. We show that pAVEC plasmids match or exceed the performance of commonly used binary vectors for transient and transgenic expression of proteins fused to fluorophores and the proximity labelling tool, TurboID. To illustrate the versatility of the pAVEC system, we demonstrate that replacing an herbicide resistance gene with a silencing suppressor gene increases protein expression in cells by an order of magnitude. We anticipate that the pAVEC system will accelerate investigations of plant molecular biology and that the modular, intentional design of these plasmids will be useful to investigate future improvements to binary plasmid design.

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selected citations
These citations are derived from selected sources.
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
0
Average
Average
Average
gold