Difference between revisions of "Part:BBa K2505001"
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This part produces AHK4, a receptor of iP(isopentenyl adenine: kind of cytokinin). This part was introduced to <i>E.coli</i>(KMI002) and sensed iP synthesized by human cell. The genes(<i>ara</i>BAD: P<sub>BAD</sub>) are transcriptionally regulated by AraC protein.  The gene(<i>ahk4</i>) is derived from <i>A.thaliana</i> and optimized for <i>E.coli</i> codon in reference to the codon usage. AHK4 protein is transmembrane protein and toxic for <i>E.coli</i> cell. Thus, We regulated expression of AHK4 by P<sub>BAD/araC</sub>: L-arabinose operon(tight transcriptionaly regulation system) .
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This part produces AHK4, a receptor of iP(isopentenyl adenine: kind of cytokinin). This part was introduced to <i>E.coli</i>(KMI002) and sensed iP synthesized by human cell. The gene(<i>ahk4</i>) is derived from <i>A.thaliana</i> and optimized for <i>E.coli</i> codon in reference to the codon usage. The AHK4 protein is transmembrane protein and toxic for <i>E.coli</i> cell. Thus, We regulated expression of AHK4 by P<sub>BAD/araC</sub>: L-arabinose operon(tight transcriptionaly regulation system).
  
 
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==Characterization==
 
==Characterization==
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We tested AHK4 protein receives iP and induces histidine kinase phosphorylation in <i>E.coli</i> cell. Our objective was to characterize the function of AHK4 protein.
  
 
===Materials & methods===
 
===Materials & methods===

Revision as of 16:35, 25 October 2017

pBad/araC-rbs-ahk4

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1205
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1294
    Illegal BamHI site found at 1144
    Illegal BamHI site found at 2375
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 979
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 961
    Illegal SapI site found at 1542
    Illegal SapI.rc site found at 3177


This part produces AHK4, a receptor of iP(isopentenyl adenine: kind of cytokinin). This part was introduced to E.coli(KMI002) and sensed iP synthesized by human cell. The gene(ahk4) is derived from A.thaliana and optimized for E.coli codon in reference to the codon usage. The AHK4 protein is transmembrane protein and toxic for E.coli cell. Thus, We regulated expression of AHK4 by PBAD/araC: L-arabinose operon(tight transcriptionaly regulation system).

Characterization

We tested AHK4 protein receives iP and induces histidine kinase phosphorylation in E.coli cell. Our objective was to characterize the function of AHK4 protein.

Materials & methods