Difference between revisions of "Part:BBa K4286107:Design"
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+ | <span class='h3bb'>Sequence and Features</span> | ||
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+ | ===Functional Parameters=== | ||
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===Source=== | ===Source=== | ||
We input the CDS sequence of PG in the professional shRNA design webpage, and the RNAi fragment is automatically designed by the program. Our examination confirmed that the sense portion of this sequence perfectly matches the gene encoding PG of Rhizoctonia solani AG1-IA. | We input the CDS sequence of PG in the professional shRNA design webpage, and the RNAi fragment is automatically designed by the program. Our examination confirmed that the sense portion of this sequence perfectly matches the gene encoding PG of Rhizoctonia solani AG1-IA. | ||
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===References=== | ===References=== |
Latest revision as of 14:42, 3 October 2023
shRNA (PG)-1
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
We have confirmed that this siRNA sequence has a good binding ability to Rhizoctonia solani. The intrinsic order of this sequence is sense RNAi fragment — loop — antisense RNAi fragment. This sequence was assembled in the pET28a (+) plasmid containing the IPTG-inducible phage T7 promoter and subsequently transferred into RNase-deficient E. coli HT115 (DE3). In our project, shRNA will be industrially produced by E. coli on a large scale, and shRNA will be purified and sprayed on rice fields to inhibit Rhizoctonia solani.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 1946
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 1046
Illegal NgoMIV site found at 1091
Illegal NgoMIV site found at 1385
Illegal NgoMIV site found at 1499
Illegal AgeI site found at 2015 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 66
Illegal BsaI site found at 1232
Illegal BsaI.rc site found at 1109
Source
We input the CDS sequence of PG in the professional shRNA design webpage, and the RNAi fragment is automatically designed by the program. Our examination confirmed that the sense portion of this sequence perfectly matches the gene encoding PG of Rhizoctonia solani AG1-IA.
References
[1]Rao, T.B., Chopperla, R., Methre, R. et al. Pectin induced transcriptome of a Rhizoctonia solani strain causing sheath blight disease in rice reveals insights on key genes and RNAi machinery for development of pathogen derived resistance. Plant Mol Biol 100, 59–71 (2019). https://doi.org/10.1007/s11103-019-00843-9 [2]Chen, X., Lili, L., Zhang, Y. et al. Functional analysis of polygalacturonase gene RsPG2 from Rhizoctonia solani, the pathogen of rice sheath blight. Eur J Plant Pathol 149, 491–502 (2017). https://doi.org/10.1007/s10658-017-1198-5