Difference between revisions of "Part:BBa K1412014"
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This part consists of a <i>[http://en.wikipedia.org/wiki/Chemotaxis CheZ]</i> gene which can express CheZ protein deciding <i>E.coli</i> whether tumble or swim straight. In this light, we can characterize the efficiency of promoter by just change different promoters. Then we can characterize the efficiency of promoter via measuring the migration distance positively associated with the expression strength of CheZ.
 
This part consists of a <i>[http://en.wikipedia.org/wiki/Chemotaxis CheZ]</i> gene which can express CheZ protein deciding <i>E.coli</i> whether tumble or swim straight. In this light, we can characterize the efficiency of promoter by just change different promoters. Then we can characterize the efficiency of promoter via measuring the migration distance positively associated with the expression strength of CheZ.
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[[File:Characterization_process_design.png|500px]]
 
[[File:Characterization_process_design.png|500px]]

Revision as of 17:17, 6 October 2014

Characterize the efficiency of pTetR (R0040) with chemotaxis

BBa_K1412014: pTetR-RBS(1.0)-CheZ-TT

This part consists of a [http://en.wikipedia.org/wiki/Chemotaxis CheZ] gene which can express CheZ protein deciding E.coli whether tumble or swim straight. In this light, we can characterize the efficiency of promoter by just change different promoters. Then we can characterize the efficiency of promoter via measuring the migration distance positively associated with the expression strength of CheZ.


Characterization process design.png


Usage

When we want to characterize the efficiency of promoter, we usually link the promoter with GFP, then characterize the promoter by just measure the fluorescence intensity of GFP. In our part, you need just link RBS(1.0) after a pTetR promoter and before a CheZ gene, ending with a TT terminator(pTetR-RBS(1.0)-CheZ-TT). Then transfer this gene circuit into E.coli (CheZ knocked out), and coat plates, culuture on semi-solid medium to measure the migration diameter of E.coli.


Relevant parts

BBa_K1412000: pLac-RBS(1.0)-CheZ-TT CheZ generator under pLac promoter

BBa_K1412005: RBS(1.0)-CheZ-TT

BBa_K1412006: RBS(0.01)-CheZ-TT

BBa_K1412007: RBS(0.3)-CheZ-TT

BBa_K1412614: pBAD-RBS(1.0)-CheZ-TT Characterize the efficiency of promoters with chemotaxis

BBa_K1412801: pLac-RBS(0.01)-CheZ-TT Characterize the efficiency of RBS with chemotaxis

BBa_K1412829: pLac-RBS(0.3)-CheZ-TT Characterize efficiency of RBS with chemotaxis


Notes

Source

BBa_R0040

BBa_B0034

BBa_K629003

BBa_B0015


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]


Protocol

Genetic link stage

1.As a skeleton, TT terminator link with CheZ.

2.Then the skeleton CheZ-TT link to RBS(1.0).

3.Next, link RBS(1.0)-CheZ-TT with promoter pTetR.

Characterization stage

1.Transfer the part pTetR-RBS(1.0)-CheZ-TT into E.coli (CheZ knock out), and coat plates, culture for hours to measure the migration diameter of E.coli.


Reference