Research output: Contribution to journal › Conference article › peer-review
In vivo dsRNA production system for silencing the phytoene desaturase gene of potato Solanum tuberosum. / Ivanov, Artemii; Golubeva, Tatiana.
In: E3S Web of Conferences, Vol. 494, 04045, 22.02.2024.Research output: Contribution to journal › Conference article › peer-review
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TY - JOUR
T1 - In vivo dsRNA production system for silencing the phytoene desaturase gene of potato Solanum tuberosum
AU - Ivanov, Artemii
AU - Golubeva, Tatiana
N1 - This work was supported by the Russian Science Foundation (grant No. 22-76-00037).
PY - 2024/2/22
Y1 - 2024/2/22
N2 - 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).
AB - 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).
UR - https://www.scopus.com/record/display.uri?eid=2-s2.0-85188238158&origin=inward&txGid=1067571b767f1b1622cac8d741546f62
UR - https://www.mendeley.com/catalogue/4ef172f5-72d5-3773-865b-e3cbffa0a789/
U2 - 10.1051/e3sconf/202449404045
DO - 10.1051/e3sconf/202449404045
M3 - Conference article
VL - 494
JO - E3S Web of Conferences
JF - E3S Web of Conferences
SN - 2555-0403
M1 - 04045
T2 - 2023 International Conference on Ensuring Sustainable Development: Ecology, Energy, Earth Science and Agriculture
Y2 - 21 December 2023 through 22 December 2023
ER -
ID: 61246106