| Name | Type | Description | Designer | Length |
|---|---|---|---|---|
| BBa_K2377000 | DNA | Intron loop with Aphids specificity | Zhicheng Lin | 194 |
| BBa_K2377001 | DNA | Intron loop with Arabidopsis specificity | He Jindian, Zhicheng Lin | 106 |
| BBa_K2377002 | Coding | Celluse coding sequence | Zhicheng Lin | 1500 |
| BBa_K2377003 | Coding | Pectinase coding sequence | Zhicheng Lin | 1047 |
| BBa_K2377010 | DNA | EcrL1 , the sense fragment of Ecr gene | He Jindian, Zhang Shuting | 523 |
| BBa_K2377011 | DNA | EcrL2 , the anti-sense fragment of Ecr gene | He Jindian, Zhang Shuting, Zhicheng Lin | 523 |
| BBa_K2377012 | DNA | Trxz sense fragment | He Jindian | 552 |
| BBa_K2377013 | DNA | Trxz anti-sense fragment | He Jindian | 552 |
| BBa_K2377014 | Regulatory | zarp, zinc two-component relulatory system | Zhou Tuoyu | 201 |
| BBa_K2377015 | DNA | loop120 | Zhicheng Lin | 121 |
| Name | Type | Description | Designer | Length |
|---|---|---|---|---|
| BBa_K2377004 | DNA | Thioredoxin Z with two reverse T7 promoters and double terminators to generate dsRNA | Zhicheng Lin, Ren Yi, He jindian, Huang Zonghui | 904 |
| BBa_K2377005 | DNA | TrxzL, A developed element to generate trxz hpRNA with a normal loop | Zhicheng Lin, Ren Yi, He jindian, Huang Zonghui | 1401 |
| BBa_K2377006 | DNA | Replace intron loop with loop120( selected form A-1 chloramphenicol O-acetyltransferase) in BBa_K2377005 | Zhicheng Lin, Ren Yi, He jindian, Huang Zonghui | 1386 |
| BBa_K2377007 | DNA | EcrL, one element to genarete Ecr hpRNA | Zhicheng Lin | 1358 |
| BBa_K2377008 | DNA | ecrN | Zhicheng Lin | 1431 |
| BBa_K2377009 | DNA | zrap ribB | Zhou Tuoyu | 1029 |
| Name | Type | Description | Designer | Length |
|---|---|---|---|---|
| BBa_K2377016 | Plasmid backbone | pDP: plasmid backbone for dsRNA production | Zhicheng Lin | 2349 |
Main part
BBa_K2377004
BBa_K2377009 is the initial part of our project. It is made up of Thioredoxin z (TRXz) gene fragment with two reverse T7 promoters and double terminators. It have been shown that silencing of TRX z result in a PEP-deficient phenotype which renders the plants incapable to grow autotrophically with their leaves being yellow.
BBa_K2377005
Traditional way to generate RNAi gene silencer using two reverse T7 promotors has a relative low efficiency due to the interference of two RNA polymerases during transcription process. To address this problem, we designed hpRNA with a loop between sense and antisense form of target sequence. It can be generated using only one promotor.
At first, we used loop120 (a 120bp sequence selected from Chl resistant gene.) as our loop. Afterwards, we saw from papers that the intron sequence in hpRNA can be spliced by spliceosome in plant. This process raises the efficiency of RNAi in plant by an unknown mechanism. Therefore, intron could be a better option of loop.
BBa_K2377007
Acyrthosiphon pisum ecdysone receptor (Ecr) is predicted by automated computational analysis. This record is derived from a genomic sequence (NW_003383721.1). ECR silencing can effectively reduce the expression of vital genes for Aphids development, thereby reducing the survival rate of insects and reproduction rate. This part is the sense arm of EcrL with loop120.
BBa_K2377002/BBa_K2377003
We design this two part for helping our dsRNA invade plant. Traces of pectinase and cellulase can cause
