Rapid Deletion Production in Fungi via <em>Agrobacterium</em> Mediated Transformation of OSCAR Deletion Constructs

Scott E. Gold; Zahi Paz; Mar&#237;a D. Garc&#237;a-Pedrajas; Anthony E. Glenn

doi:10.3791/55239

Journal

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Genetics

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

Rapid Deletion Production in Fungi via <em>Agrobacterium</em> Mediated Transformation of OSCAR Deletion Constructs

JoVE Journal
Genetics

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JoVE Journal Genetics

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

10,012 Views

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11:24 min

•

June 12, 2017

DOI:

10.3791/55239-v

DOI:

10.3791/55239-v

•

11:24 min

June 12, 2017

•

10006 Views

Scott E. Gold¹, Zahi Paz², María D. García-Pedrajas³, Anthony E. Glenn¹

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

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.