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

DOI:

11:24 min

June 12, 2017

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Capítulos

  • 00:05Título
  • 01:02Primer Design for PCR Amplification of Gene Flanks
  • 02:49Production of OSCAR Constructs
  • 05:48Agrobacterium tumefaciens Mediated Transformation (ATMT) of Fungi
  • 07:45Deletion Mutant Identification by PCR
  • 08:53Results: The OSCAR Method Generates a Gene Deletion Plasmid for Fungi in a Single Reaction
  • 10:23Conclusion

Summary

Tadução automática

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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