Difference between revisions of "Part:BBa K1455002"
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<partinfo>BBa_K1455002 short</partinfo> | <partinfo>BBa_K1455002 short</partinfo> | ||
| − | This device is used to express LuxO D47E by arabinose induction | + | This device is used to express LuxO D47E by arabinose induction and at the same time to measure the expression of LuxOD 47E in an indirectly way by GFP fluoresence. |
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LuxOD 47E acts like a phosphorilated LuxO, which is an essential protein of ''Vibrio spp.'' quorum sensing system. LuxO is the final protein in a phophorilation cascade and binds, as a trancription facotr, to pqrr promoters. LuxOD 47E is able to activate the genes downstream of a pqrr promoter. | LuxOD 47E acts like a phosphorilated LuxO, which is an essential protein of ''Vibrio spp.'' quorum sensing system. LuxO is the final protein in a phophorilation cascade and binds, as a trancription facotr, to pqrr promoters. LuxOD 47E is able to activate the genes downstream of a pqrr promoter. | ||
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| + | Arabinose inducible promoter used here is a strong one. | ||
The following pictures were taken using a Eclipse-Ti Nikon® ECLIPSE Ti inverted microscope equiped with an ANDOR iXon Ultra 897 camera. The filter used was FITC. | The following pictures were taken using a Eclipse-Ti Nikon® ECLIPSE Ti inverted microscope equiped with an ANDOR iXon Ultra 897 camera. The filter used was FITC. | ||
| − | Here is ''E. coli'' DH5α carrying this device after one hour of induction with 0.1 % arabinose | + | Here is ''E. coli'' DH5α carrying this device after one hour of induction with 0.1 % arabinose, light (left) and FITC (right): |
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| + | [[File:unahoraraluz.png|350px]] | ||
| + | [[File:unahoraarafitc.png|350px]] | ||
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| − | + | And after 3 hours (both with FITC): | |
| + | [[File:treshorasara.png|350px]] | ||
| + | [[File:treshorasara2.png|350px]] | ||
Latest revision as of 02:18, 8 October 2014
pBAD - luxO D47E - GFP
This device is used to express LuxO D47E by arabinose induction and at the same time to measure the expression of LuxOD 47E in an indirectly way by GFP fluoresence.
LuxOD 47E acts like a phosphorilated LuxO, which is an essential protein of Vibrio spp. quorum sensing system. LuxO is the final protein in a phophorilation cascade and binds, as a trancription facotr, to pqrr promoters. LuxOD 47E is able to activate the genes downstream of a pqrr promoter.
Arabinose inducible promoter used here is a strong one.
The following pictures were taken using a Eclipse-Ti Nikon® ECLIPSE Ti inverted microscope equiped with an ANDOR iXon Ultra 897 camera. The filter used was FITC.
Here is E. coli DH5α carrying this device after one hour of induction with 0.1 % arabinose, light (left) and FITC (right):
And after 3 hours (both with FITC):
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 125
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 65
Illegal XhoI site found at 292 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 980
Illegal BsaI.rc site found at 2189
Illegal SapI.rc site found at 942