Difference between revisions of "Part:BBa K4226000"
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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. | 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. | ||
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=Usage and Biology= | =Usage and Biology= | ||
| − | + | ==Characterized by CAFA_China 2022== | |
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*We designed the genetic circuit that contains sulAp ([[Part:BBa_K518010|sulA promoter]]), Amp30E, relE gene ([[Part:BBa_K185047|RelE toxin]]), mScarlet-I ([[Part:BBa_K3977002|mSarlet-I]]) and 3WJ-Bro, and transferred it into DH10B competent cells (The control group did not contain the Amp30E Amplification Device). | *We designed the genetic circuit that contains sulAp ([[Part:BBa_K518010|sulA promoter]]), Amp30E, relE gene ([[Part:BBa_K185047|RelE toxin]]), mScarlet-I ([[Part:BBa_K3977002|mSarlet-I]]) and 3WJ-Bro, and transferred it into DH10B competent cells (The control group did not contain the 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. <br> | *After bacterial culture and UV induction, we measured the OD600 and fluorescence values under 485/510nm. Then, the fluorescence per OD was calculated. <br> | ||
Revision as of 06:46, 26 September 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.
Usage and Biology
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 the 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.
- The curve of experimental group was significantly higher than that of control group and the amount of relE RNA synthesis gradually increased with incubation time. According to the results, the 3WJ-Bro can be used to ligate fluorescent aptamers to examine the relE RNA synthesis at transcriptional level.
