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Part:BBa_K4604026
piG_K12BSa (LacIprom_riboK12_lacI_trcProm_sfGFP)
piG_K12BSa consists of the LacI promoter, the AdoCbl riboswitch from E. coli K12, lacI and the trc promoter. The trc promoter can be induced with IPTG. This BioBrick is a sensor for AdoCbl (a bioavailable form of B12) with a variable marker gene.
Characterization
To verify the performance of the BioBrick we added different concentrations of AdoCbl to the cultures and tested the sensitivity of the sensor by fluorescent measurement. AdoCbl is taken up by the cells, binds to the riboswitch which repressed LacI production thus enabling the expression of sfGFP.
Fluorescent measurement, as shown in figure 2, indicates that at a concentration of 100 nM AdoCbl, the riboswitch is saturated and can no longer recognize an increase of AdoCbl. This data verifies the sensibility of the sensor; it can detect AdoCbl in different concentrations and accordingly regulate the gene downstream of it. However, we also noted a high background fluorescence without the addition of AdoCbl.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 215
- 1000COMPATIBLE WITH RFC[1000]
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