Part:BBa_K1682016
IPTG induced RFP reporter system
HKUST-RICE team designed an IPTG inducible RFP reporter system. This was achieved by combining the constitutive promoter BBa_J23101 with LacI generator BBa_P0412, and a RFP coding device (BBa_J04450) driven by Plac.
Mechanism
Figure 1. Schematic diagram of the inducible Plac-mRFP construct. In the absence of Isopropyl b-D-1-thiogalactopyranoside (IPTG), LacI represses Plac promoter. When IPTG is present, LacI protein is released from the <i>lacO operator, triggering the production of mRFP.
Usage and Functionality
In order to check the functionality of the construct for use in sensing IPTG concentrations, characterisation was done inducing the cell containing the BBa_K1682016 construct in pSB3K3 under varing IPTG concentrations.
Figure 2. Dose response of E. coli DH10B containing pSB3K3-BBa_K1682016 to varing IPTG levels. After overnight induction of cell culture incubated in M9-glycerol at 37˚C, measurements for population averaged assay were taken by fluorometry.
The results from characterisation show that the working concentration of the constructed IPTG sensor is at around 0.001 to 0.5 mM IPTG, with an 4.5 fold dynamic range.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1084
Illegal NheI site found at 1107 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 2285
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 781
Illegal AgeI site found at 893 - 1000COMPATIBLE WITH RFC[1000]
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