Difference between revisions of "Part:BBa K3885353"
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Under the P28 promoter, the Tet operator tetO is expressed along with the production of green fluorescent protein.The transcription rate of the P28 promoter can be detected in response to MGapt. | Under the P28 promoter, the Tet operator tetO is expressed along with the production of green fluorescent protein.The transcription rate of the P28 promoter can be detected in response to MGapt. | ||
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===Usage and Biology=== | ===Usage and Biology=== | ||
| − | + | Malachite green aptamer (MGA), an RNA aptamer that can specifically bind to malachite green (MG) dye and light up the fluorescence more than 2000-fold compared to negligible fluorescence of free MG. The MGA-MG system can be used for the quantitative analysis of RNA.<br> | |
| + | In Cell-Free systems, the strength of the P28 promoter has not been quantified. We cloned the MG aptamer on the 3’ end of P28-tetO-deGFP. Therefore, the transcription rate of P28 can be estimated. The optimal Ex/Em wavelengths are 625/655nm. The dye should be used under 20 μM, otherwise it is toxic to the reaction.<br> | ||
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<span class='h3bb'>Sequence and Features</span> | <span class='h3bb'>Sequence and Features</span> | ||
Latest revision as of 03:50, 20 October 2021
P28-tetO-degfp-MGapt
Under the P28 promoter, the Tet operator tetO is expressed along with the production of green fluorescent protein.The transcription rate of the P28 promoter can be detected in response to MGapt.
Usage and Biology
Malachite green aptamer (MGA), an RNA aptamer that can specifically bind to malachite green (MG) dye and light up the fluorescence more than 2000-fold compared to negligible fluorescence of free MG. The MGA-MG system can be used for the quantitative analysis of RNA.
In Cell-Free systems, the strength of the P28 promoter has not been quantified. We cloned the MG aptamer on the 3’ end of P28-tetO-deGFP. Therefore, the transcription rate of P28 can be estimated. The optimal Ex/Em wavelengths are 625/655nm. The dye should be used under 20 μM, otherwise it is toxic to the reaction.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 763
Illegal BamHI site found at 795 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]