Difference between revisions of "Part:BBa K3815012"
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__NOTOC__ | __NOTOC__ | ||
| − | <partinfo>BBa_K3815012 short</partinfo> | + | <partinfo>BBa_K3815012 short</partinfo><br> |
| − | This is a section of | + | This is a section of the TrxZ gene of Arabidopsis thaliana to synthesize dsRNA for RNAi. To product dsRNA, this sequence is inserted in the L4440 plasmid, and transformed into HT115(DE3). |
<h3><font size="3"></font> purpose </h3> | <h3><font size="3"></font> purpose </h3> | ||
| − | Trx-Z is a gene responsible for chloroplast formation during early development in Arabidopsis thaliana. Silencing this | + | Trx-Z is a gene responsible for chloroplast formation during early development in Arabidopsis thaliana. Silencing this gene will whiten leafs of Arabidopsis thaliana[1]. |
<br> | <br> | ||
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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> | 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<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 | + | 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.<br><br> |
HT115(DE3)<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> | 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> | ||
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<partinfo>BBa_K3815012 SequenceAndFeatures</partinfo> | <partinfo>BBa_K3815012 SequenceAndFeatures</partinfo> | ||
<h3><font size="3"> cloning </font> </h3> | <h3><font size="3"> cloning </font> </h3> | ||
| − | This part was inserted in L4440. The L4440 has two t7 promoters, and | + | This part was inserted in L4440. The L4440 has two t7 promoters, and these parts are transcribed from both sides. |
<br> | <br> | ||
| + | ==Reference== | ||
| + | 1 Chen, Z., He, J., Luo, P., Li, X., and Gao, Y. (2018). Production of functional double-stranded RNA using a prokaryotic expression system in Escherichia coli. Microbiologyopen e787. | ||
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Latest revision as of 02:20, 22 October 2021
Part of the TrxZ gene of A. thaliana to synthesize dsRNA for RNAi
This is a section of the TrxZ gene of Arabidopsis thaliana to synthesize dsRNA for RNAi. To product dsRNA, this sequence is inserted in the L4440 plasmid, and transformed into HT115(DE3).
purpose
Trx-Z is a gene responsible for chloroplast formation during early development in Arabidopsis thaliana. Silencing this gene will whiten leafs of Arabidopsis thaliana[1].
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 are annealed.
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]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 442
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
cloning
This part was inserted in L4440. The L4440 has two t7 promoters, and these parts are transcribed from both sides.
Reference
1 Chen, Z., He, J., Luo, P., Li, X., and Gao, Y. (2018). Production of functional double-stranded RNA using a prokaryotic expression system in Escherichia coli. Microbiologyopen e787.