Difference between revisions of "Part:BBa K1412000"

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Figure 2. Comparision with [https://parts.igem.org/Part:BBa_J04450 BBa_J04450] and BBa_K1412000
 
Figure 2. Comparision with [https://parts.igem.org/Part:BBa_J04450 BBa_J04450] and BBa_K1412000
  
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BBa_J04450 and BBa_K1412000 are transformed into <i>CL-1</i> strains ( with <i>CheZ</i> gene knocked out of genmo) respectively. From the result, we can see that the chemotaxis diameter of BBa_K1412000 (d2) is longer than BBa_J04450 (d1), which means BBa_K1412000 could work as expectation.
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<i>CheZ</i> and <i>LacI</i> are both knocked out of the genome of <i>CL-1</i>, so the <i>CL-1</i> does not have the ability of chemotaxis and will not inhibit the expression of Lac promoter. Therefore <i>CheZ</i> in [https://parts.igem.org/Part:BBa_K1412000  BBa_K1412000 ] can be expressed correctly, then the ability of chemotaxis recovers. While without <i>CheZ</i>, [https://parts.igem.org/Part:BBa_J04450 BBa_J04450]  can't migrate. All of the above are reflected on the difference in the diameter of chemotaxis: d2 > d1.
  
 
We use M63 semi-solid plate to observe the behavior of the two <i>E.coli(CL-1)</i> strain. We dot 3μL of the two bacterias on the same plate, then place the plate at 37℃. By comparing the chemotaxis diameter of the bacteria on the plate, we conclude BBa_K1412000 has the ability of chemotaxis.
 
We use M63 semi-solid plate to observe the behavior of the two <i>E.coli(CL-1)</i> strain. We dot 3μL of the two bacterias on the same plate, then place the plate at 37℃. By comparing the chemotaxis diameter of the bacteria on the plate, we conclude BBa_K1412000 has the ability of chemotaxis.

Revision as of 12:47, 11 October 2014

CheZ generator under Lac promoter


This part is consist of Lac promoter and CheZ generator.

CheZ can dephosphorylate CheY to make the flagellar motor rotate counterclockwise so that the cell to run and tumble.Without CheZ, CheY is phosphorylated, which can bind to the flagellar motor protein FliM, causing the cell to tumble.so the cell tumbles only. The expression of CheZ allows E.coli to navigate gradients of natural chemoattractants.

At the same time, we know that Lac promoter is an inverting regulator sensitive to LacI and CAP, while LacI can be inhibited by IPTG. So with LacI protein, CAP protein and IPTG,we can adjust the expression of CheZ via Lac promoter to control the movement of E.coli. File:PLac-RBS-CheZ-TT.png

Usage and Biology

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]

Experimental Data

CL-1 and BBa K1412000.png

Figure 2. Comparision with BBa_J04450 and BBa_K1412000

CheZ and LacI are both knocked out of the genome of CL-1, so the CL-1 does not have the ability of chemotaxis and will not inhibit the expression of Lac promoter. Therefore CheZ in BBa_K1412000 can be expressed correctly, then the ability of chemotaxis recovers. While without CheZ, BBa_J04450 can't migrate. All of the above are reflected on the difference in the diameter of chemotaxis: d2 > d1.

We use M63 semi-solid plate to observe the behavior of the two E.coli(CL-1) strain. We dot 3μL of the two bacterias on the same plate, then place the plate at 37℃. By comparing the chemotaxis diameter of the bacteria on the plate, we conclude BBa_K1412000 has the ability of chemotaxis.