Cargando…

BioVector, a flexible system for gene specific-expression in plants

BACKGROUND: Functional genomic research always needs to assemble different DNA fragments into a binary vector, so as to express genes with different tags from various promoters with different levels. The cloning systems available bear similar disadvantages, such as promoters/tags are fixed on a bina...

Descripción completa

Detalles Bibliográficos
Autores principales: Wang, Xu, Fan, Chengming, Zhang, Xiaomei, Zhu, Jinlong, Fu, Yong-Fu
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4235170/
https://www.ncbi.nlm.nih.gov/pubmed/24304941
http://dx.doi.org/10.1186/1471-2229-13-198
_version_ 1782344979079757824
author Wang, Xu
Fan, Chengming
Zhang, Xiaomei
Zhu, Jinlong
Fu, Yong-Fu
author_facet Wang, Xu
Fan, Chengming
Zhang, Xiaomei
Zhu, Jinlong
Fu, Yong-Fu
author_sort Wang, Xu
collection PubMed
description BACKGROUND: Functional genomic research always needs to assemble different DNA fragments into a binary vector, so as to express genes with different tags from various promoters with different levels. The cloning systems available bear similar disadvantages, such as promoters/tags are fixed on a binary vector, which is generally with low cloning efficiency and limited for cloning sites if a novel promoter/tag is in need. Therefore, it is difficult both to assemble a gene and a promoter together and to modify the vectors in hand. Another disadvantage is that a long spacer from recombination sites, which may be detrimental to the protein function, exists between a gene and a tag. Multiple GATEWAY system only resolves former problem at the expense of very low efficiency and expensive for multiple LR reaction. RESULTS: To improve efficiency and flexibility for constructing expression vectors, we developed a platform, BioVector, by combining classical restriction enzyme/ligase strategy with modern Gateway DNA recombination system. This system included a series of vectors for gene cloning, promoter cloning, and binary vector construction to meet various needs for plant functional genomic study. CONCLUSION: This BioVector platform makes it easy to construct any vectors to express a target gene from a specific promoter with desired intensity, and it is also waiting to be freely modified by researchers themselves for ongoing demands. This idea can also be transferred to the different fields including animal or yeast study.
format Online
Article
Text
id pubmed-4235170
institution National Center for Biotechnology Information
language English
publishDate 2013
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-42351702014-11-19 BioVector, a flexible system for gene specific-expression in plants Wang, Xu Fan, Chengming Zhang, Xiaomei Zhu, Jinlong Fu, Yong-Fu BMC Plant Biol Methodology Article BACKGROUND: Functional genomic research always needs to assemble different DNA fragments into a binary vector, so as to express genes with different tags from various promoters with different levels. The cloning systems available bear similar disadvantages, such as promoters/tags are fixed on a binary vector, which is generally with low cloning efficiency and limited for cloning sites if a novel promoter/tag is in need. Therefore, it is difficult both to assemble a gene and a promoter together and to modify the vectors in hand. Another disadvantage is that a long spacer from recombination sites, which may be detrimental to the protein function, exists between a gene and a tag. Multiple GATEWAY system only resolves former problem at the expense of very low efficiency and expensive for multiple LR reaction. RESULTS: To improve efficiency and flexibility for constructing expression vectors, we developed a platform, BioVector, by combining classical restriction enzyme/ligase strategy with modern Gateway DNA recombination system. This system included a series of vectors for gene cloning, promoter cloning, and binary vector construction to meet various needs for plant functional genomic study. CONCLUSION: This BioVector platform makes it easy to construct any vectors to express a target gene from a specific promoter with desired intensity, and it is also waiting to be freely modified by researchers themselves for ongoing demands. This idea can also be transferred to the different fields including animal or yeast study. BioMed Central 2013-12-05 /pmc/articles/PMC4235170/ /pubmed/24304941 http://dx.doi.org/10.1186/1471-2229-13-198 Text en Copyright © 2013 Wang et al.; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Methodology Article
Wang, Xu
Fan, Chengming
Zhang, Xiaomei
Zhu, Jinlong
Fu, Yong-Fu
BioVector, a flexible system for gene specific-expression in plants
title BioVector, a flexible system for gene specific-expression in plants
title_full BioVector, a flexible system for gene specific-expression in plants
title_fullStr BioVector, a flexible system for gene specific-expression in plants
title_full_unstemmed BioVector, a flexible system for gene specific-expression in plants
title_short BioVector, a flexible system for gene specific-expression in plants
title_sort biovector, a flexible system for gene specific-expression in plants
topic Methodology Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4235170/
https://www.ncbi.nlm.nih.gov/pubmed/24304941
http://dx.doi.org/10.1186/1471-2229-13-198
work_keys_str_mv AT wangxu biovectoraflexiblesystemforgenespecificexpressioninplants
AT fanchengming biovectoraflexiblesystemforgenespecificexpressioninplants
AT zhangxiaomei biovectoraflexiblesystemforgenespecificexpressioninplants
AT zhujinlong biovectoraflexiblesystemforgenespecificexpressioninplants
AT fuyongfu biovectoraflexiblesystemforgenespecificexpressioninplants