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Construction of Artificial miRNAs to Prevent Drought Stress in Solanum tuberosum

  • Anna Wyrzykowska
  • Marcin Pieczynski
  • Zofia Szweykowska-KulinskaEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1398)

Abstract

The use of artificial microRNAs (amiRNAs) is still a relatively new technique in molecular biology with a wide range of applications in life sciences. Here, we describe the silencing of the CBP80/ABH1 gene in Solanum tuberosum with the use of amiRNA. The CBP80/ABH1 protein is part of the Cap Binding Complex (CBC), which is involved in plant responses to drought stress conditions. Transformed plants with a decreased level of CBP80/ABH1 display increased tolerance to water shortage conditions. We describe how to design amiRNA with the Web MicroRNA Designer platform in detail. Additionally, we explain how to perform all steps of a procedure aiming to obtain transgenic potato plants with the use of designed amiRNA, through callus tissue regeneration and Agrobacterium tumefaciens strain LBA4404 as a transgene carrier.

Key words

Artificial microRNA (amiRNA) Solanum tuberosum Drought stress CBP80/ABH1 Transformation Agrobacterium tumefaciens 

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Anna Wyrzykowska
    • 1
  • Marcin Pieczynski
    • 1
  • Zofia Szweykowska-Kulinska
    • 1
    Email author
  1. 1.Department of Gene Expression, Faculty of BiologyAdam Mickiewicz UniversityPoznanPoland

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