Part:BBa_K2379011
Adaptor with RNA pH meter
This construct consists of the adaptor BBa_K2379008(BBa_K2379008) and is under the influence of the pLac promoter(BBa_R0011). In addition to the adaptor, the RNA pHmeter BBa_K2379012(BBa_K2379012) is used to regulate the transcription of the genes downstream. Following the adaptor is the reporter protein GFP (BBa_E0040) and the double terminator(BBa_B0014). Since it is regulated by the RNA pHmeter, the circuit will get activated and GFP will be produced only at pH 8.5
Usage and Biology
ABOUT THIS PART:
This part is used to convert translational regulation to transcriptional regulation by means of the adaptor ( BBa_K2379008 ) . This variant of the part contains the pH Sensitive SraF gene which will control the transcription of downstream genes. This part works as expected and when GFP is placed downstream of the pLac promoter, transcription and translation of downstream genes were induced at pH 8.5
CHARACTERISATION STUDIES FOR THIS PART:
1. The above graph indicates the fluorescence expression at three different time periods (each recorded at an interval of 60 minutes)
2. The cell cultures were grown at two different pH : 7.0 and 8.5. Fluorescence values were recorded at excitation and emission peaks of 460nm and 515nm respectively.
3. Normalised fluorescence was calculated by dividing observed fluorescence by cell density at OD600.
4. Duplets were used for characterisation and the error bars indicate the Standard Deviation of the mean values.
CONCLUSION:
The results obtained for the part having adaptor combined with RNA pHmeter is as expected. There was an average 25.46% increase in the expression of GFP from pH 7 to pH 8.5 at the two time points which is certainly higher than that obtained for RNA pHmeter without adaptor. Thus, this part works as expected and shows maximum expression at pH 8.5.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 181
Illegal BsaI.rc site found at 1219
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