Difference between revisions of "Part:BBa K1682017"
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<partinfo>BBa_K1682017 short</partinfo> | <partinfo>BBa_K1682017 short</partinfo> | ||
− | This composite part is consists of a L- | + | This composite part is consists of a L-arabinose inducible <i>P<sub>araBAD</sub></i> with GFP reporter <partinfo>BBa_I13540</partinfo>, and an IPTG inducible P<sub>lac</sub> with RFP reporter [https://parts.igem.org/Part:BBa_K1682016 Part:BBa_K1682016]. |
− | The part was designed to investigate the | + | The part was designed to investigate the possible effect of parallel sensing within a single plasmid. |
==Mechanism== | ==Mechanism== | ||
[[File:Parallel_expression_BBa_K1682017.jpg|700px|centre|]] | [[File:Parallel_expression_BBa_K1682017.jpg|700px|centre|]] | ||
− | '''Figure 1. Schematic diagram of the inducible P<sub>araBAD</sub>-GFP-P<sub>lac</sub>- | + | '''Figure 1. Schematic diagram of the inducible <i>P<sub>araBAD</sub>-GFP-P<sub>lac</sub>-mrfp</i> construct.''' The system can sense L-arabinose to give GFP output or sense IPTG to give RFP output. |
==Usage and Biology== | ==Usage and Biology== | ||
− | In order to investigate the effect of parallel sensing, the dose response P<sub>araBAD</sub>- | + | In order to investigate the effect of parallel sensing, the dose response of <i>P<sub>araBAD</sub>-gfp</i> in pSB3K3-<partinfo>BBa_K1682017</partinfo> (double construct) was compared with that in pSB3K3-<partinfo>BBa_I13540</partinfo> (single construct) in <i>E. coli</i> DH10B. |
+ | |||
+ | [[File:Parallel_expression_BBa_K1682017_fig2.jpg|800px|centre|]] | ||
+ | '''Figure 2. Response curve of <i>P<sub>araBAD</sub>-gfp</i> in a single construct and double construct under varying L-arabinose concentration.'''The different blue curves indicate different induction level of the co-expressed <i>P<sub>lac</sub>-mrfp</i> construct. At varying the induction level of the co-expressed <i>P<sub>lac</sub>-mrfp</i> construct seems to have an effect on the expression level of <i>P<sub>araBAD</sub>-gfp</i> in a double construct. At a higher IPTG concentration, the maximum expression of <i>P<sub>araBAD</sub>-gfp</i> decreases. | ||
Latest revision as of 03:53, 19 September 2015
ParaBAD-GFP-Plac-RFP
This composite part is consists of a L-arabinose inducible ParaBAD with GFP reporter BBa_I13540, and an IPTG inducible Plac with RFP reporter Part:BBa_K1682016. The part was designed to investigate the possible effect of parallel sensing within a single plasmid.
Mechanism
Figure 1. Schematic diagram of the inducible ParaBAD-GFP-Plac-mrfp construct. The system can sense L-arabinose to give GFP output or sense IPTG to give RFP output.
Usage and Biology
In order to investigate the effect of parallel sensing, the dose response of ParaBAD-gfp in pSB3K3-BBa_K1682017 (double construct) was compared with that in pSB3K3-BBa_I13540 (single construct) in E. coli DH10B.
Figure 2. Response curve of ParaBAD-gfp in a single construct and double construct under varying L-arabinose concentration.The different blue curves indicate different induction level of the co-expressed Plac-mrfp construct. At varying the induction level of the co-expressed Plac-mrfp construct seems to have an effect on the expression level of ParaBAD-gfp in a double construct. At a higher IPTG concentration, the maximum expression of ParaBAD-gfp decreases.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1205
Illegal NheI site found at 3185
Illegal NheI site found at 3208 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 4386
Illegal BamHI site found at 1144 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 979
Illegal AgeI site found at 2882
Illegal AgeI site found at 2994 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1880
Illegal SapI site found at 961