| Issue |
E3S Web Conf.
Volume 494, 2024
International Conference on Ensuring Sustainable Development: Ecology, Energy, Earth Science and Agriculture (AEES2023)
|
|
|---|---|---|
| Article Number | 04045 | |
| Number of page(s) | 5 | |
| Section | Current Agricultural Development | |
| DOI | https://doi.org/10.1051/e3sconf/202449404045 | |
| Published online | 22 February 2024 | |
In vivo dsRNA production system for silencing the phytoene desaturase gene of potato Solanum tuberosum
1 Institute of Cytology and Genetics, 630090, Novosibirsk, Russia
2 Novosibirsk State University, 630090, Novosibirsk, Russia
* Corresponding author: frolova@bionet.nsc.ru
In this work, for the first time, an in vivo system was developed for the production of exogenous double-stranded RNA (dsRNA), complementary to the potato phytoene desaturase (pds) gene. For this purpose, a recombinant plasmid was constructed based on the L4440 vector; the pds gene fragment was inserted between two late T7 promoters, which allows the production of dsRNA. The resulting plasmid was used to transform the E. coli strain HT115, which is deficient in RNase III. This strain prevents the hydrolysis of the dsRNA produced in it. The presence of the required dsRNA fragments was confirmed by electrophoretic analysis. This system can be used as a positive control in experiments on regulating the activity of potato genes using spray-induced gene silencing (SIGS).
© The Authors, published by EDP Sciences, 2024
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