Coding

Part:BBa_K422007

Designed by: Katharina Zwicky   Group: iGEM10_ETHZ_Basel   (2010-10-25)

CheY (AarI A-part)

Biological Background

The chemotactic network consists of membrane (methyl accepting chemotaxis proteins: MCPs) and intracellular proteins (Che) as signal transducers and receptors. MCPs sense a minimal difference in input concentration and transduce this information unit to the repsective proteins CheW and CheA, which located inside the cell. The autophosphorylation of CheA mediated by the MCPs is the key step in tumbling induction in response to increased repellent or decreased attractant concentration. The methylation state of the MCPs is influenced by the methyltransferase CheR (transfers a methylgroup to a protein) and the demethylase CheB (cleaves a methyl group off a protein). CheA phosphorylates CheY which then diffuses through the cytoplasm to the flagellar motor protein FliM which as a response induces tumbling. The phosphatase CheZ regulates the signal termination via dephosphorylation of CheYp (the p stands for the phosphorylated form of CheY).

Cloning strategy

BBF RFC28: A method for combinatorial multi-part assembly based on the Type IIs restriction enzyme AarI. CheY-A is an A-part compatible for N-Terminal fusion. For more information see [http://2010.igem.org/Team:ETHZ_Basel/Biology/Cloning].

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 275
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 275
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 275
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 275
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
Parameters
None