Difference between revisions of "Part:BBa K3718001"
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<p>When the nanobody binds the target, dimerization of the protein CadC domains occurs. As a result, this allows the binding of the pCadBA operon region, and transcription downstream is promoted at the same time. </p> | <p>When the nanobody binds the target, dimerization of the protein CadC domains occurs. As a result, this allows the binding of the pCadBA operon region, and transcription downstream is promoted at the same time. </p> | ||
| − | + | https://2021.igem.org/wiki/images/thumb/6/63/T--Hong_Kong_UCCKE--MRP.png/800px-T--Hong_Kong_UCCKE--MRP.png | |
<p>The VHH region has high flexible possibilities as it can be customised for binding with different targets, so that any specific targets with single-domain antibodies available are potentially ligand candidates for this receptor. </p> | <p>The VHH region has high flexible possibilities as it can be customised for binding with different targets, so that any specific targets with single-domain antibodies available are potentially ligand candidates for this receptor. </p> | ||
Latest revision as of 10:44, 15 October 2021
Transmembrane receptor with Customizable VHH region
Our Modular Receptor Platform (MRP) is designed according to the paper published in 2017. The modular biosensor is composed of the E. coli CadC DNA binding domain, a juxtamembrane bridge region, an artificial 16-Leucine transmembrane region, and a transmembrane receptor with customizable VHH region, which is a restriction site.
When the nanobody binds the target, dimerization of the protein CadC domains occurs. As a result, this allows the binding of the pCadBA operon region, and transcription downstream is promoted at the same time.
The VHH region has high flexible possibilities as it can be customised for binding with different targets, so that any specific targets with single-domain antibodies available are potentially ligand candidates for this receptor.
Users can add their own antibody VHH region after the transmembrane region DNA via restriction ligation. The KasI restriction site, together with sequence GCC, is added at the C-terminus of the transmembrane region. The addition of the restriction site is 3 bp. Therefore, the coding region, which includes the user’s VHH, is in-frame. A minimum addition of nucleotides is added in order to reduce any effect of the additional codon to the coding region. At the same time, KasI recognizes a 6bp site, so that the chances of having the cut site in other regions of the user's DNA are relatively low.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal prefix found in sequence at 1
Illegal suffix found in sequence at 563 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 1
Illegal SpeI site found at 564
Illegal PstI site found at 578
Illegal NotI site found at 7
Illegal NotI site found at 571 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 1
- 23INCOMPATIBLE WITH RFC[23]Illegal prefix found in sequence at 1
Illegal suffix found in sequence at 564 - 25INCOMPATIBLE WITH RFC[25]Illegal prefix found in sequence at 1
Illegal XbaI site found at 16
Illegal SpeI site found at 564
Illegal PstI site found at 578 - 1000COMPATIBLE WITH RFC[1000]