Reporter
Part:BBa_K4226000
Designed by: Siyu Han Group: iGEM22_CAFA_China (2022-09-14)
Revision as of 06:27, 26 September 2022 by Tigress K (Talk | contribs) (→Characterized by CAFA_China 2022)
3WJ-Bro
3WJ-Bro is used as a protein- independent reporter system based on a fluorescent RNA aptamer. It is applied as a gene marker in creating a system for reporting the presence and expression of target genes. The 3WJ-Bro can be used to ligate fluorescent aptamers to examine the relE RNA at transcriptional level in Escherichia coli cells, obviating the need for accumulation of foreign proteins.
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Characterized by CAFA_China 2022
- We designed the genetic circuit that contains sulAp (sulA promoter), Amp30E, relE gene (RelE toxin), mScarlet-I (mSarlet-I) and 3WJ-Bro, and transferred it into DH10B competent cells (The control group did not contain Amp30E Amplification Device).
- After bacterial culture and UV induction, we measured the OD600 and fluorescence values under 485/510nm. Then, the fluorescence per OD was calculated.
- With the lengthening of culture time, the RNA synthesis of relE increased gradually in the experimental group. According to the results, the 3WJ-Bro can be used to ligate fluorescent aptamers to examine the relE RNA synthesis at transcriptional level.
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Categories
Parameters
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