Difference between revisions of "Part:BBa K1682016"
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<partinfo>BBa_K1682016 short</partinfo> | <partinfo>BBa_K1682016 short</partinfo> | ||
| − | HKUST-RICE team designed an IPTG inducible RFP reporter system. | + | HKUST-RICE team designed an IPTG inducible RFP reporter system. This was achieved by combining the constitutive promoter <partinfo>BBa_J23101</partinfo> with LacI generator <partinfo>BBa_P0412</partinfo>, and a RFP coding device (<partinfo>BBa_J04450</partinfo>) driven by Plac. |
==Mechanism== | ==Mechanism== | ||
| − | [[File:HKUST-RICE BBa K1682016.1.jpg|500px|]] | + | [[File:HKUST-RICE BBa K1682016.1.jpg|500px|centre|]] |
'''Figure 1. Schematic diagram of the inducible Plac-mRFP construct.''' '''(a)'''In the absence of Isopropyl b-D-1-thiogalactopyranoside (IPTG), LacI represses Plac. '''(b)'''When IPTG is present, LacI protein is repressed, triggering the production of mRFP. | '''Figure 1. Schematic diagram of the inducible Plac-mRFP construct.''' '''(a)'''In the absence of Isopropyl b-D-1-thiogalactopyranoside (IPTG), LacI represses Plac. '''(b)'''When IPTG is present, LacI protein is repressed, triggering the production of mRFP. | ||
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==Usage and Functionality== | ==Usage and Functionality== | ||
| − | In order to check the functionality of the construct for use in sensing IPTG concentrations. | + | In order to check the functionality of the construct for use in sensing IPTG concentrations, characterisation was done inducing the cell containing the <partinfo>BBa_K1682016</partinfo> construct in pSB3K3 under varing IPTG concentrations. |
| − | + | [[File:HKUST-RICE BBa K1682016 fig2.jpg|500px|centre|]] | |
| + | '''Figure 2. Dose response of <i>E. coli</i> DH10B containing pSB3K3-BBa_K1682016 to varing IPTG levels.''' Measurements were taken after overnight induction of cell culture incubated in M9-glycerol at 37degC. | ||
| + | |||
| + | 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. | ||
| + | |||
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| + | ==Sequence and Features== | ||
<partinfo>BBa_K1682016 SequenceAndFeatures</partinfo> | <partinfo>BBa_K1682016 SequenceAndFeatures</partinfo> | ||
Revision as of 09:18, 15 September 2015
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. (a)In the absence of Isopropyl b-D-1-thiogalactopyranoside (IPTG), LacI represses Plac. (b)When IPTG is present, LacI protein is repressed, 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. Measurements were taken after overnight induction of cell culture incubated in M9-glycerol at 37degC.
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]