GreenGate---a novel, versatile, and efficient cloning system for plant transgenesis.

Building expression constructs for transgenesis is one of the fundamental day-to-day tasks in modern biology. Traditionally it is based on a multitude of type II restriction endonucleases and T4 DNA ligase. Especially in case of long inserts and applications requiring high-throughput, this approach...

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Autores principales:	Athanasios Lampropoulos, Zoran Sutikovic, Christian Wenzl, Ira Maegele, Jan U Lohmann, Joachim Forner
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2013
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/d90544c3c9654bdfbc4ec3623a9551e4
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spelling	oai:doaj.org-article:d90544c3c9654bdfbc4ec3623a9551e42021-11-18T08:40:56ZGreenGate---a novel, versatile, and efficient cloning system for plant transgenesis.1932-620310.1371/journal.pone.0083043https://doaj.org/article/d90544c3c9654bdfbc4ec3623a9551e42013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24376629/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203Building expression constructs for transgenesis is one of the fundamental day-to-day tasks in modern biology. Traditionally it is based on a multitude of type II restriction endonucleases and T4 DNA ligase. Especially in case of long inserts and applications requiring high-throughput, this approach is limited by the number of available unique restriction sites and the need for designing individual cloning strategies for each project. Several alternative cloning systems have been developed in recent years to overcome these issues, including the type IIS enzyme based Golden Gate technique. Here we introduce our GreenGate system for rapidly assembling plant transformation constructs, which is based on the Golden Gate method. GreenGate cloning is simple and efficient since it uses only one type IIS restriction endonuclease, depends on only six types of insert modules (plant promoter, N-terminal tag, coding sequence, C-terminal tag, plant terminator and plant resistance cassette), but at the same time allows assembling several expression cassettes in one binary destination vector from a collection of pre-cloned building blocks. The system is cheap and reliable and when combined with a library of modules considerably speeds up cloning and transgene stacking for plant transformation.Athanasios LampropoulosZoran SutikovicChristian WenzlIra MaegeleJan U LohmannJoachim FornerPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 8, Iss 12, p e83043 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Athanasios Lampropoulos Zoran Sutikovic Christian Wenzl Ira Maegele Jan U Lohmann Joachim Forner GreenGate---a novel, versatile, and efficient cloning system for plant transgenesis.
description	Building expression constructs for transgenesis is one of the fundamental day-to-day tasks in modern biology. Traditionally it is based on a multitude of type II restriction endonucleases and T4 DNA ligase. Especially in case of long inserts and applications requiring high-throughput, this approach is limited by the number of available unique restriction sites and the need for designing individual cloning strategies for each project. Several alternative cloning systems have been developed in recent years to overcome these issues, including the type IIS enzyme based Golden Gate technique. Here we introduce our GreenGate system for rapidly assembling plant transformation constructs, which is based on the Golden Gate method. GreenGate cloning is simple and efficient since it uses only one type IIS restriction endonuclease, depends on only six types of insert modules (plant promoter, N-terminal tag, coding sequence, C-terminal tag, plant terminator and plant resistance cassette), but at the same time allows assembling several expression cassettes in one binary destination vector from a collection of pre-cloned building blocks. The system is cheap and reliable and when combined with a library of modules considerably speeds up cloning and transgene stacking for plant transformation.
format	article
author	Athanasios Lampropoulos Zoran Sutikovic Christian Wenzl Ira Maegele Jan U Lohmann Joachim Forner
author_facet	Athanasios Lampropoulos Zoran Sutikovic Christian Wenzl Ira Maegele Jan U Lohmann Joachim Forner
author_sort	Athanasios Lampropoulos
title	GreenGate---a novel, versatile, and efficient cloning system for plant transgenesis.
title_short	GreenGate---a novel, versatile, and efficient cloning system for plant transgenesis.
title_full	GreenGate---a novel, versatile, and efficient cloning system for plant transgenesis.
title_fullStr	GreenGate---a novel, versatile, and efficient cloning system for plant transgenesis.
title_full_unstemmed	GreenGate---a novel, versatile, and efficient cloning system for plant transgenesis.
title_sort	greengate---a novel, versatile, and efficient cloning system for plant transgenesis.
publisher	Public Library of Science (PLoS)
publishDate	2013
url	https://doaj.org/article/d90544c3c9654bdfbc4ec3623a9551e4
work_keys_str_mv	AT athanasioslampropoulos greengateanovelversatileandefficientcloningsystemforplanttransgenesis AT zoransutikovic greengateanovelversatileandefficientcloningsystemforplanttransgenesis AT christianwenzl greengateanovelversatileandefficientcloningsystemforplanttransgenesis AT iramaegele greengateanovelversatileandefficientcloningsystemforplanttransgenesis AT janulohmann greengateanovelversatileandefficientcloningsystemforplanttransgenesis AT joachimforner greengateanovelversatileandefficientcloningsystemforplanttransgenesis
_version_	1718421472761872384

GreenGate---a novel, versatile, and efficient cloning system for plant transgenesis.

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