Cargando…

The Bxb1 recombination system demonstrates heritable transmission of site-specific excision in Arabidopsis

BACKGROUND: The mycobacteriophage large serine recombinase Bxb1 catalyzes site-specific recombination between its corresponding attP and attB recognition sites. Previously, we and others have shown that Bxb1 has catalytic activity in various eukaryotic species including Nicotiana tabacum, Schizosacc...

Descripción completa

Detalles Bibliográficos
Autores principales: Thomson, James G, Chan, Ronald, Smith, Jamison, Thilmony, Roger, Yau, Yuan-Yeu, Wang, YueJu, Ow, David W
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3341217/
https://www.ncbi.nlm.nih.gov/pubmed/22436504
http://dx.doi.org/10.1186/1472-6750-12-9
_version_ 1782231503689744384
author Thomson, James G
Chan, Ronald
Smith, Jamison
Thilmony, Roger
Yau, Yuan-Yeu
Wang, YueJu
Ow, David W
author_facet Thomson, James G
Chan, Ronald
Smith, Jamison
Thilmony, Roger
Yau, Yuan-Yeu
Wang, YueJu
Ow, David W
author_sort Thomson, James G
collection PubMed
description BACKGROUND: The mycobacteriophage large serine recombinase Bxb1 catalyzes site-specific recombination between its corresponding attP and attB recognition sites. Previously, we and others have shown that Bxb1 has catalytic activity in various eukaryotic species including Nicotiana tabacum, Schizosaccharomyces pombe, insects and mammalian cells. RESULTS: In this work, the Bxb1 recombinase gene was transformed and constitutively expressed in Arabidopsis thaliana plants harboring a chromosomally integrated attP and attB-flanked target sequence. The Bxb1 recombinase successfully excised the target sequence in a conservative manner and the resulting recombination event was heritably transmitted to subsequent generations in the absence of the recombinase transgene. In addition, we also show that Bxb1 recombinase expressing plants can be manually crossed with att-flanked target transgenic plants to generate excised progeny. CONCLUSION: The Bxb1 large serine recombinase performs site-specific recombination in Arabidopsis thaliana germinal tissue, producing stable lines free of unwanted DNA. The precise site-specific deletion produced by Bxb1 in planta demonstrates that this enzyme can be a useful tool for the genetic engineering of plants without selectable marker transgenes or other undesirable exogenous sequences.
format Online
Article
Text
id pubmed-3341217
institution National Center for Biotechnology Information
language English
publishDate 2012
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-33412172012-05-02 The Bxb1 recombination system demonstrates heritable transmission of site-specific excision in Arabidopsis Thomson, James G Chan, Ronald Smith, Jamison Thilmony, Roger Yau, Yuan-Yeu Wang, YueJu Ow, David W BMC Biotechnol Research Article BACKGROUND: The mycobacteriophage large serine recombinase Bxb1 catalyzes site-specific recombination between its corresponding attP and attB recognition sites. Previously, we and others have shown that Bxb1 has catalytic activity in various eukaryotic species including Nicotiana tabacum, Schizosaccharomyces pombe, insects and mammalian cells. RESULTS: In this work, the Bxb1 recombinase gene was transformed and constitutively expressed in Arabidopsis thaliana plants harboring a chromosomally integrated attP and attB-flanked target sequence. The Bxb1 recombinase successfully excised the target sequence in a conservative manner and the resulting recombination event was heritably transmitted to subsequent generations in the absence of the recombinase transgene. In addition, we also show that Bxb1 recombinase expressing plants can be manually crossed with att-flanked target transgenic plants to generate excised progeny. CONCLUSION: The Bxb1 large serine recombinase performs site-specific recombination in Arabidopsis thaliana germinal tissue, producing stable lines free of unwanted DNA. The precise site-specific deletion produced by Bxb1 in planta demonstrates that this enzyme can be a useful tool for the genetic engineering of plants without selectable marker transgenes or other undesirable exogenous sequences. BioMed Central 2012-03-21 /pmc/articles/PMC3341217/ /pubmed/22436504 http://dx.doi.org/10.1186/1472-6750-12-9 Text en Copyright ©2012 Thomson 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 Research Article
Thomson, James G
Chan, Ronald
Smith, Jamison
Thilmony, Roger
Yau, Yuan-Yeu
Wang, YueJu
Ow, David W
The Bxb1 recombination system demonstrates heritable transmission of site-specific excision in Arabidopsis
title The Bxb1 recombination system demonstrates heritable transmission of site-specific excision in Arabidopsis
title_full The Bxb1 recombination system demonstrates heritable transmission of site-specific excision in Arabidopsis
title_fullStr The Bxb1 recombination system demonstrates heritable transmission of site-specific excision in Arabidopsis
title_full_unstemmed The Bxb1 recombination system demonstrates heritable transmission of site-specific excision in Arabidopsis
title_short The Bxb1 recombination system demonstrates heritable transmission of site-specific excision in Arabidopsis
title_sort bxb1 recombination system demonstrates heritable transmission of site-specific excision in arabidopsis
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3341217/
https://www.ncbi.nlm.nih.gov/pubmed/22436504
http://dx.doi.org/10.1186/1472-6750-12-9
work_keys_str_mv AT thomsonjamesg thebxb1recombinationsystemdemonstratesheritabletransmissionofsitespecificexcisioninarabidopsis
AT chanronald thebxb1recombinationsystemdemonstratesheritabletransmissionofsitespecificexcisioninarabidopsis
AT smithjamison thebxb1recombinationsystemdemonstratesheritabletransmissionofsitespecificexcisioninarabidopsis
AT thilmonyroger thebxb1recombinationsystemdemonstratesheritabletransmissionofsitespecificexcisioninarabidopsis
AT yauyuanyeu thebxb1recombinationsystemdemonstratesheritabletransmissionofsitespecificexcisioninarabidopsis
AT wangyueju thebxb1recombinationsystemdemonstratesheritabletransmissionofsitespecificexcisioninarabidopsis
AT owdavidw thebxb1recombinationsystemdemonstratesheritabletransmissionofsitespecificexcisioninarabidopsis
AT thomsonjamesg bxb1recombinationsystemdemonstratesheritabletransmissionofsitespecificexcisioninarabidopsis
AT chanronald bxb1recombinationsystemdemonstratesheritabletransmissionofsitespecificexcisioninarabidopsis
AT smithjamison bxb1recombinationsystemdemonstratesheritabletransmissionofsitespecificexcisioninarabidopsis
AT thilmonyroger bxb1recombinationsystemdemonstratesheritabletransmissionofsitespecificexcisioninarabidopsis
AT yauyuanyeu bxb1recombinationsystemdemonstratesheritabletransmissionofsitespecificexcisioninarabidopsis
AT wangyueju bxb1recombinationsystemdemonstratesheritabletransmissionofsitespecificexcisioninarabidopsis
AT owdavidw bxb1recombinationsystemdemonstratesheritabletransmissionofsitespecificexcisioninarabidopsis