Chapter title |
Construction of Artificial miRNAs to Prevent Drought Stress in Solanum tuberosum.
|
---|---|
Chapter number | 21 |
Book title |
Environmental Responses in Plants
|
Published in |
Methods in molecular biology, January 2016
|
DOI | 10.1007/978-1-4939-3356-3_21 |
Pubmed ID | |
Book ISBNs |
978-1-4939-3354-9, 978-1-4939-3356-3
|
Authors |
Anna Wyrzykowska, Marcin Pieczynski, Zofia Szweykowska-Kulinska |
Editors |
Paula Duque |
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. |
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