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United States Department of Agriculture

Agricultural Research Service

Research Project: POSITIONAL CLONING IN MAIZE OF GENES THAT REGULATE PLANT ARCHITECTURE

Location: Plant Gene Expression Center Albany_CA

Title: From gateway to multisite gateway in one recombination event

Authors
item Magnani, Esteban - ARS-UCB PLNT GENE EXP CTR
item Bartling, Linnea - ARS-UCB PLNT GENE EXP CTR
item Hake, Sarah

Submitted to: BioMed Central (BMC) Molecular Biology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: December 6, 2006
Publication Date: December 6, 2006
Repository URL: http://www.biomedcentral.com/content/pdf/1471-2199-7-46.pdf
Citation: Magnani, E., Bartling, L., Hake, S.C. 2006. From gateway to multisite gateway in one recombination event. BMC Microbiology. 7(1):46.

Interpretive Summary: Molecular biology allows one to combine different pieces of DNA for study and analysis. Invitrogen Gateway technology exploits the integrase/att site-specific recombination system for directional cloning of PCR products and the subsequent subcloning into destination vectors. We created a construct to easily convert any available Gateway destination vector to a MultiSite Gateway vector in a single recombination reaction. Our tool makes MultiSite Gateway a more widely accessible technology and expands its applications by exploiting all the features of the Gateway vectors already available.

Technical Abstract: We created a construct, pDONR-R4-R3, to easily convert any available Gateway destination vector to a MultiSite Gateway vector in a single recombination reaction. In addition, we designed pDONR-R4-R3 so that DNA fragments already cloned upstream or downstream of the Gateway cassette in the original destination vectors can still be utilized for promoter-gene or translational fusions after the conversion.

Last Modified: 7/24/2014
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