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Rapid Deletion Production in Fungi via Agrobacterium Mediated Transformation of OSCAR Deletion Constructs
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Rapid Deletion Production in Fungi via Agrobacterium Mediated Transformation of OSCAR Deletion Constructs

Rapid Deletion Production in Fungi via Agrobacterium Mediated Transformation of OSCAR Deletion Constructs

9,645 Views

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11:24 min

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June 12, 2017

DOI:

10.3791/55239-v

DOI:
10.3791/55239-v

•

11:24 min
June 12, 2017

•

1 Views
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1Toxicology and Mycotoxin Research Unit,NPRC, USDA-ARS, 2Hazera Seeds LTD, Brurim, 3Instituto de Hortofruticultura Subtropical y Mediterránea “La Mayora” – Universidad de Málaga – Consejo Superior de Investigaciones Científicas (IHSM-UMA-CSIC), Estación Experimental “La Mayora”

Summary

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Gene deletion mutants generated through homologous recombination are the gold standard for gene function studies. The OSCAR (One Step Construction of Agrobacterium-Recombination-ready-plasmids) method for rapid generation of deletion constructs is described. Agrobacterium mediated fungal transformation follows. Finally, a PCR based confirmation method of gene deletions in fungal transformants is presented.

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Rapid Deletion ProductionFungiAgrobacteriumOSCAR Deletion ConstructsHomologous RecombinationFunctional GeneticsGene Deletion MutantsDelsGate SystemPrimer DesignPCRDeletion Confirmation

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Cite this Article

Gold, S. E., Paz, Z., García-Pedrajas, M. D., Glenn, A. E. Rapid Deletion Production in Fungi via Agrobacterium Mediated Transformation of OSCAR Deletion Constructs. J. Vis. Exp. (124), e55239, doi:10.3791/55239 (2017).

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