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Plants 2017, 6(4), 55; doi:10.3390/plants6040055

Development of pGEMINI, a Plant Gateway Destination Vector Allowing the Simultaneous Integration of Two cDNA via a Single LR-Clonase Reaction

School of Biological Sciences, Wivenhoe Park, University of Essex, Colchester CO4 3SQ, UK
Genetics, Genomics and Breeding, NIAB EMR, New Road, East Malling, Kent ME19 6BJ, UK
Author to whom correspondence should be addressed.
Received: 27 October 2017 / Revised: 6 November 2017 / Accepted: 10 November 2017 / Published: 12 November 2017
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Gateway technology has been used to facilitate the generation of a large number of constructs for the modification of plants for research purposes. However, many of the currently available vectors only allow the integration of a single cDNA of interest into an expression clone. The ability to over-express multiple genes in combination is essential for the study of plant development where several transcripts have a role to play in one or more metabolic processes. The tools to carry out such studies are limited, and in many cases rely on the incorporation of cDNA into expression systems via conventional cloning, which can be both time consuming and laborious. To our knowledge, this study reports on the first development of a vector allowing the simultaneous integration of two independent cDNAs via a single LR-clonase reaction. This vector “pGEMINI” represents a powerful molecular tool offering the ability to study the role of multi-cDNA constructs on plant development, and opens up the process of gene stacking and the study of gene combinations through transient or stable transformation procedures. View Full-Text
Keywords: gateway; gene stacking; LR clonase; pGEMINI; fluorescence gateway; gene stacking; LR clonase; pGEMINI; fluorescence

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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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Exposito-Rodriguez, M.; Laissue, P.P.; López-Calcagno, P.E.; Mullineaux, P.M.; Raines, C.A.; Simkin, A.J. Development of pGEMINI, a Plant Gateway Destination Vector Allowing the Simultaneous Integration of Two cDNA via a Single LR-Clonase Reaction. Plants 2017, 6, 55.

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