Difference between revisions of "Part:BBa K629003"
 
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<partinfo>BBa_K629003 short</partinfo>
 
<partinfo>BBa_K629003 short</partinfo>
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<b>Usage and Biology</b><br>
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== Background ==
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Chemotaxis is the cell’s system for directed movement which allows the cell to move towards nutrition sources as well as away from toxic substances. The chemotactic system is a complex chain of phosphorylation reactions. At the end of the chain is the flagellum-binding protein CheY. The phosphorylated CheY (CheY-p) binds the flagella and causes the cell to tumble in place, essentially remaining immotile. The phosphatase CheZ dephosphorylates CheY, causing CheY to unbind to the flagella, resulting in directed movement of the cell [1].
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<b>References</b><br>
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Polar localization is dependent on CheA-short. CheZ is a cytosolic phosphatase which functions in the chemotaxis signal transduction complex.
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[1] <i>Madigan MT, Martinko JM, Stahl DA, Clark DP. 2010. Brock biology of microorganisms. 13th ed. Benjamin Cummings, San Francisco.</i><br>
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Plays an important role in bacterial chemotaxis signal transduction pathway by accelerating the dephosphorylation of phosphorylated CheY (CheY-P).
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== Expression in ''E. Coli'' ==
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Before we started our functional test of CheZ, we have utilized western blot to determine whether there are enough soluble proteins in E. Coli. With 6×His tag on pET-32a, the result shows that CheZ has more expression with induction of IPTG.
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Note: Because the promoter is Plac here, the expression has a background, which means control also expresses some cheZ without induction of IPTG.
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[[Image:CheZ.jpg]]
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== Picture of Chemotaxis in ''cheZ'' knocked-out strain ==
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The main purpose of cheZ is to control chemotaxis in E. Coli. What we have done is to use different promoter to test how to get the better result from expression of CheZ. Below are pictures and parameters of chemotaxis test.
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Nalidixic acid Induction of promoter ''recAp''. If you need higher resolution pictures, please feel free to cantact us.
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[[Image:ddH2O.jpg]]
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[[Image:5 NAL.jpg]]
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[[Image:10 NAL.jpg]]
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===Usage and Biology===
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>

Latest revision as of 17:07, 15 October 2014

cheZ, chemotaxis regulator, protein phosphatase for CheY

Usage and Biology
Chemotaxis is the cell’s system for directed movement which allows the cell to move towards nutrition sources as well as away from toxic substances. The chemotactic system is a complex chain of phosphorylation reactions. At the end of the chain is the flagellum-binding protein CheY. The phosphorylated CheY (CheY-p) binds the flagella and causes the cell to tumble in place, essentially remaining immotile. The phosphatase CheZ dephosphorylates CheY, causing CheY to unbind to the flagella, resulting in directed movement of the cell [1].

References
[1] Madigan MT, Martinko JM, Stahl DA, Clark DP. 2010. Brock biology of microorganisms. 13th ed. Benjamin Cummings, San Francisco.
Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]