Difference between revisions of "Part:BBa K3071000"
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<partinfo>BBa_K3071000 short</partinfo> | <partinfo>BBa_K3071000 short</partinfo> | ||
===Description=== | ===Description=== | ||
| − | This the protein-encoding gene for the Sensory/regulatory protein RpfC, which is codon-optimized to express in <i>escherica coli</i> K12 strain. This part is developed and improved based on the previous record part ([https://parts.igem.org/Part:BBa_K1315002 BBa K1315002]). The western blot data of our composite part | + | This the protein-encoding gene for the Sensory/regulatory protein RpfC, which is codon-optimized to express in <i>escherica coli</i> K12 strain. This part is developed and improved based on the previous record part ([https://parts.igem.org/Part:BBa_K1315002 BBa K1315002]). The western blot data of our composite part ([https://parts.igem.org/Part:BBa_K3071018 BBa_K3071018]) shows it could be correctly expressed in <i>E. coli</i>. |
===Biology=== | ===Biology=== | ||
RpfC is a kinase located in the cell membrane of bacteria acting as a sensor. It consists of three domains, including the input domain, histidine kinase (HisKA)domain, CheY-like two-component receiver domain (REC) and the C-terminal histidine phosphotransfer (HPT) domain. The sensory domain is composed of 5 transmembrane helices with periplasmic and cytoplasmic loops less than 20 amino acids long. | RpfC is a kinase located in the cell membrane of bacteria acting as a sensor. It consists of three domains, including the input domain, histidine kinase (HisKA)domain, CheY-like two-component receiver domain (REC) and the C-terminal histidine phosphotransfer (HPT) domain. The sensory domain is composed of 5 transmembrane helices with periplasmic and cytoplasmic loops less than 20 amino acids long. | ||
Revision as of 16:06, 18 October 2019
Sensory/regulatory protein RpfC from Xanthomononas campestris pv. campestris
Description
This the protein-encoding gene for the Sensory/regulatory protein RpfC, which is codon-optimized to express in escherica coli K12 strain. This part is developed and improved based on the previous record part (BBa K1315002). The western blot data of our composite part (BBa_K3071018) shows it could be correctly expressed in E. coli.
Biology
RpfC is a kinase located in the cell membrane of bacteria acting as a sensor. It consists of three domains, including the input domain, histidine kinase (HisKA)domain, CheY-like two-component receiver domain (REC) and the C-terminal histidine phosphotransfer (HPT) domain. The sensory domain is composed of 5 transmembrane helices with periplasmic and cytoplasmic loops less than 20 amino acids long.
Usage
RpfC is mainly used as a receptor for diffusible signal factor (DSF) in cell-cell communication. When the DSF signal is received, active autophosphorylation starts at the His198 of HisKA domain. It will then be followed by a phosphorelay through the Asp512 and His657 in the REC domain and the HPT domain respectively. The phosphotransfer lastly activates the RpfG by phosphorylation at the REC domain of RpfG.(figure 1)
Besides activating the RpfG signal protein, the second role of RpfC is to interact with RpfF, a DSF synthase, by its REC domain, in order to suppress the DSF production in that bacterial cell.
Characterization
Research showed that point mutations at His 198, Asp512 or His657 resulted in the reduction of virulence factors, extracellular cellulases, and proteases, demonstrating the importance of these three amino acids in the process of RpfC-RpfG signaling and the role of RpfC in virulence factor production.
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 303
Illegal AgeI site found at 1135
Illegal AgeI site found at 1614
Illegal AgeI site found at 1698 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 352
Illegal BsaI.rc site found at 216