Part:BBa_K1864000
CSBV-RdRP
Our part is the gene of CSBV's RNA dependent RNA polymerase(RdRp). We transferred the gene of RdRp into plasmid L4440 ,and transformed the recombinant plasmid into E.coli strain HT115 to express double stranded RNA of RdRp.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 837
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 110
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Application: To produce double stranded RNA of RNA dependent RNA polymerase which is able to initiate the process of RNA-interference
Group: FAFU-CHINA
Author: Ruicheng Dai & Changlong Lu
Summary: We used this part to produce dsRNA of RdRp, which is essential in our project to silence the RdRp gene of Sacbrood virus（CSBV）, preventing it from producing a protein.
Design
In our project,we have been aiming to control Chinese Sacbrood virus through silence of CSBV’s RdRp (RNA-dependent RNA polymerase) gene by using RNA-interference technology. We built recombinant L4440 plasmid containing dual T7 promoter and the gene of RNA Dependent RNA polymerase, and we transformed it to HT115 strain. After the enlarge cultivation of HT115, we added the engineered bacteria to the forage of honeybees. To test the effect of dsRdRp, we then did a infectious experiment by feeding infected hives with dsRdRp, and collected data to study the pupation rate of infected hives under different concentrations.
We expected this part to express homologous double-stranded RNA of RdRp. And the result of infectious experiment indicated that this part works as expected.
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