Amplification of genome-integrated BeYDV replicons by transient expression of Rep in Arabidopsis thaliana

Bean yellow dwarf geminivirus (BeYDV) has been used as a potential vector to improve foreign gene expression, specifically, to achieve higher yields of vaccine proteins in plants. Previously, we have shown that when the BeYDV replication initiator protein Rep was provided in trans, replication and g...

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Autores principales:	Moon,Yong-Sun, Hefferon,Kathleen L
Lenguaje:	English
Publicado:	Pontificia Universidad Católica de Valparaíso 2007
Materias:	Agrobacterium-mediated expression BeYDV-based vector gene amplification GUS expression Rep expression transgenic Arabidopsis
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582007000400001
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spelling	oai:scielo:S0717-345820070004000012009-01-19Amplification of genome-integrated BeYDV replicons by transient expression of Rep in Arabidopsis thalianaMoon,Yong-SunHefferon,Kathleen L Agrobacterium-mediated expression BeYDV-based vector gene amplification GUS expression Rep expression transgenic Arabidopsis Bean yellow dwarf geminivirus (BeYDV) has been used as a potential vector to improve foreign gene expression, specifically, to achieve higher yields of vaccine proteins in plants. Previously, we have shown that when the BeYDV replication initiator protein Rep was provided in trans, replication and gene expression of GUS were enhanced enormously from a BeYDV expression vector in a transient assay system. In this paper, transgenic lines of Arabidopsis (cv. Columbia) were generated harboring the BeYDV cis-acting elements required for replication. Constructs encoding BeYDV Rep or intronless Rep open reading frames (ORFs) were transiently introduced into transgenic plants via Agrobacterium-mediated infiltration in order to examine the relative levels of replication and expression of the genome-integrated GUS reporter gene. This study shows that expression of Rep protein was regulated in trans from a separate cassette which enabled the rescue, replication and enhancement of the genome-integrated GUS gene in transgenic Arabidopsis. We conclude that Rep expression can be effectively controlled in Arabidopsis plants, and that regulation of Rep expression can result in the amplification of a genome-integrated foreign gene by circumventing the negative effects of gene silencing.info:eu-repo/semantics/openAccessPontificia Universidad Católica de ValparaísoElectronic Journal of Biotechnology v.10 n.4 20072007-10-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582007000400001en10.4067/S0717-34582007000400001
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	Agrobacterium-mediated expression BeYDV-based vector gene amplification GUS expression Rep expression transgenic Arabidopsis
spellingShingle	Agrobacterium-mediated expression BeYDV-based vector gene amplification GUS expression Rep expression transgenic Arabidopsis Moon,Yong-Sun Hefferon,Kathleen L Amplification of genome-integrated BeYDV replicons by transient expression of Rep in Arabidopsis thaliana
description	Bean yellow dwarf geminivirus (BeYDV) has been used as a potential vector to improve foreign gene expression, specifically, to achieve higher yields of vaccine proteins in plants. Previously, we have shown that when the BeYDV replication initiator protein Rep was provided in trans, replication and gene expression of GUS were enhanced enormously from a BeYDV expression vector in a transient assay system. In this paper, transgenic lines of Arabidopsis (cv. Columbia) were generated harboring the BeYDV cis-acting elements required for replication. Constructs encoding BeYDV Rep or intronless Rep open reading frames (ORFs) were transiently introduced into transgenic plants via Agrobacterium-mediated infiltration in order to examine the relative levels of replication and expression of the genome-integrated GUS reporter gene. This study shows that expression of Rep protein was regulated in trans from a separate cassette which enabled the rescue, replication and enhancement of the genome-integrated GUS gene in transgenic Arabidopsis. We conclude that Rep expression can be effectively controlled in Arabidopsis plants, and that regulation of Rep expression can result in the amplification of a genome-integrated foreign gene by circumventing the negative effects of gene silencing.
author	Moon,Yong-Sun Hefferon,Kathleen L
author_facet	Moon,Yong-Sun Hefferon,Kathleen L
author_sort	Moon,Yong-Sun
title	Amplification of genome-integrated BeYDV replicons by transient expression of Rep in Arabidopsis thaliana
title_short	Amplification of genome-integrated BeYDV replicons by transient expression of Rep in Arabidopsis thaliana
title_full	Amplification of genome-integrated BeYDV replicons by transient expression of Rep in Arabidopsis thaliana
title_fullStr	Amplification of genome-integrated BeYDV replicons by transient expression of Rep in Arabidopsis thaliana
title_full_unstemmed	Amplification of genome-integrated BeYDV replicons by transient expression of Rep in Arabidopsis thaliana
title_sort	amplification of genome-integrated beydv replicons by transient expression of rep in arabidopsis thaliana
publisher	Pontificia Universidad Católica de Valparaíso
publishDate	2007
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582007000400001
work_keys_str_mv	AT moonyongsun amplificationofgenomeintegratedbeydvrepliconsbytransientexpressionofrepinarabidopsisthaliana AT hefferonkathleenl amplificationofgenomeintegratedbeydvrepliconsbytransientexpressionofrepinarabidopsisthaliana
_version_	1718441773269778432

Amplification of genome-integrated BeYDV replicons by transient expression of Rep in Arabidopsis thaliana

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