Difference between revisions of "Part:BBa K3815013"
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__NOTOC__ | __NOTOC__ | ||
<partinfo>BBa_K3815013 short</partinfo> | <partinfo>BBa_K3815013 short</partinfo> | ||
− | This is a section of EPH1 gene of Ipomoea nil to synthesize dsRNA for RNAi. To product dsRNA this sequence inserted in L4440 plasmid, and transformed into HT115(DE3). | + | This is a section of EPH1 gene of Ipomoea nil to synthesize dsRNA for RNAi. To product dsRNA, this sequence is inserted in L4440 plasmid, and transformed into HT115(DE3). |
<h3><font size="3"></font> purpose </h3> | <h3><font size="3"></font> purpose </h3> | ||
− | This is a gene for a transcription factor that positively regulates PCD in Ipomoea nil. | + | 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=== | ===Usage and Biology=== | ||
+ | RNAi<br> | ||
+ | RNAi (RNA interference) is a process in which externally introduced dsRNA suppresses the expression of genes that have complementary sequences to the dsRNA.<br><br> | ||
+ | L4440<br> | ||
+ | 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 anneal.<br><br> | ||
+ | HT115(DE3)<br> | ||
+ | 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.<br><br> | ||
+ | |||
+ | <h3><font size="3"></font>Sequence and features </h3> | ||
− | |||
<partinfo>BBa_K3815013 SequenceAndFeatures</partinfo> | <partinfo>BBa_K3815013 SequenceAndFeatures</partinfo> | ||
+ | |||
+ | <h3><font size="3"> cloning </font> </h3> | ||
+ | This part was inserted in L4440. The L4440 has two t7 promoters, and this part are transcribed from both sides. | ||
Revision as of 10:49, 21 October 2021
Part of the EPH1 gene of Ipomoea nil to synthesize dsRNA for RNAi
This is a section of EPH1 gene of Ipomoea nil to synthesize dsRNA for RNAi. To product dsRNA, this sequence is inserted in L4440 plasmid, and transformed into HT115(DE3).
purpose
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
RNAi (RNA interference) is a process in which externally introduced dsRNA suppresses the expression of genes that have complementary sequences to the dsRNA.
L4440
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 anneal.
HT115(DE3)
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[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 9
- 1000COMPATIBLE WITH RFC[1000]
cloning
This part was inserted in L4440. The L4440 has two t7 promoters, and this part are transcribed from both sides.