Part:BBa_K4208001
PctD EnvZ HAMP Hybrid
Usage
This basic part has been developed by team Tuebingen 2022 [1] to sense acetylcholine and choline (and potentially Anatoxin A) with a Hybrid receptor encoding for the PctD Sensing Domain and the EnvZ HAMP and Histidine Kinase/Phosphatase domain.
Biology
PctD is a scarcely characterized chemoreceptor, found in Pseudomonas aeruginosa. It has been found that it binds to choline and acetylcholine. [2] Anatoxin A mimics acetylcholine and binds to the nicotinic Acetylcholine receptors. 10.1016/j.scitotenv.2020.139515 To our knowledge, Anatoxin A has not yet been shown to bind PctD.
Hybrid receptors combining sensing domain from chemoreceptors and two-component systems has been a field of research since the late 1980s. The common ancestry and a shared structure for these two sensing systems have allowed to mix up sensing domains together with
Results
In our project, we have successful cloned this receptor in the pKG116 backbone plasmid that allows the expression of proteins under the regulation of the pnahG promoter [3] that we have induced with acetylsalycilic acid.
In qualitative and quantitative experiments with fluorescence microscopy and a plate reader experiment we show fluorescence of the E. coli strain containg both this hybrid receptor in the BBa_K4208004 composite part and the BBa_K4208006 reporter plasmid.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 40
Illegal NgoMIV site found at 94
Illegal NgoMIV site found at 844
Illegal NgoMIV site found at 1090 - 1000COMPATIBLE WITH RFC[1000]
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