Part:BBa_K1333204
Or2-RBS-GFP-terminator
This is a reporter which combines plac Or2-62 and the generator of GFP. In the beginning of the Or2 sequence, there is a binding site for protein lambda cI called lambda cI operator. In Bacterial Two-hybrid System, the protein lambda cI is linked to a molecular called bait. And the corresponding target is linked to rpoA, which is the α subunit of RNA polymerase. The Or2 is reponsible for the recruitment of lambda cI-bait. When the bait interacts with the target, the rpoA is carried to downstream promoter and activates the downstream expression which is GFP in this case. Stronger interactions between baits and targets indicate higher concentration of rpoA near the promoter and higher level of transcription of GFP. Hence, we can tell the interactional degree between baits and targets from the fluorescence intensity of GFP.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 741
Functional Parameters
BBa_K1333204 is the most important part in the Bacterial Two-Hybrid System. And its effectiveness was tested in the whole Bacterial Two-Hybrid System as the following experiments. It is on the backbone of pSB4A5. The plasmid is shown here.
| color | fluorescent green |
//function/reporter/fluorescence
| color | fluorescent green |