Part of the EPH1 gene of Ipomoea nil to synthesize dsRNA for RNAi
This is a section of the EPH1 gene of Ipomoea nil to synthesize dsRNA for RNAi. To product dsRNA, this sequence is inserted in the L4440 plasmid, and transformed into HT115(DE3).
This is a gene for a transcription factor that positively regulates PCD in Ipomoea nil. Silencing this gene is expected to extend the flowering time of Ipomoea nil.
Usage and Biology
RNAi (RNA interference) is a process in which externally introduced dsRNA suppresses the expression of genes that have complementary sequences to the dsRNA.
L4440 is a plasmid vector having two convergent T7 promoters adjacent to the multi-cloning site. By inserting a portion of the target gene sequence into the multi-cloning site of this plasmid, the target sequence is transcribed from both sides, and dsRNA can be obtained when both parts are annealed.
HT115 (DE3) is an RNase III-deficient E. coli strain that has been modified to express T7 RNA polymerase from an IPTG-inducible promoter. It lacks dsRNA-specific RNase III, which allows it to produce high levels of specific dsRNA. These attributes allow HT115 (DE3) to be a promising strain for the preparation of large amounts of viral dsRNA in vivo.
Sequence and features
- 10COMPATIBLE WITH RFC
- 12COMPATIBLE WITH RFC
- 21COMPATIBLE WITH RFC
- 23COMPATIBLE WITH RFC
- 25Illegal NgoMIV site found at 9
- 1000COMPATIBLE WITH RFC
This part was inserted in L4440. The L4440 has two t7 promoters, and this part is transcribed from both sides.
1 Shibuya, K., Shimizu, K., Niki, T., and Ichimura, K. (2014). Identification of a NAC transcription factor, EPHEMERAL1, that controls petal senescence in Japanese morning glory. Plant J. 79, 1044–1051.