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Rapid Deletion Production in Fungi via Agrobacterium Mediated Transformation of OSCAR Deletion Constructs
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遗传学
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JoVE 杂志 遗传学
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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