Difference between revisions of "Part:BBa K1412829"
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<bbpart>BBa_K1412000</bbpart>
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BBa_K1412000: pLac-RBS(1.0)-CheZ-TT CheZ generator under pLac promoter
  
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<bbpart>BBa_K1412005</bbpart>
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BBa_K1412005: RBS(1.0)-CheZ-TT
  
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<bbpart>BBa_K1412006</bbpart>
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BBa_K1412006: RBS(0.01)-CheZ-TT
  
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<bbpart>BBa_K1412007</bbpart>
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BBa_K1412007: RBS(0.3)-CheZ-TT
  
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<bbpart>BBa_K1412014</bbpart>
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BBa_K1412014: pTet-RBS(1.0)-CheZ-TT Characterize the efficiency of promoters with chemotaxis
  
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<bbpart>BBa_K1412614</bbpart>
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BBa_K1412614: pBAD-RBS(1.0)-CheZ-TT Characterize the efficiency of promoters with chemotaxis
  
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<bbpart>BBa_K1412801</bbpart>
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BBa_K1412801: pLac-RBS(0.01)-CheZ-TT Characterize the efficiency of RBS with chemotaxis
  
  

Revision as of 16:44, 6 October 2014

Characterize efficiency of RBS with chemotaxis

BBa_K1412829: Plac-RBS(0.3)-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 RBS or promoter efficiency by just change different promoters or ribosome binding sites(RBS). Then we can characterize the efficiency of RBS and 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 RBS, we usually link the RBS with GFP, and then characterize the RBS by just measure the fluorescence intensity of GFP. In our part, you only need to link RBS after a Plac promoter and before a CheZ gene, ending with a TT terminator(Plac-RBS(changeable)-CheZ-TT). Then transfer this gene circuit into E.coli CL-1 (CheZ knocked out), and coat plates, culture 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_K1412014: pTet-RBS(1.0)-CheZ-TT Characterize the efficiency of promoters with chemotaxis

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


Notes

Source

BBa_R0010

BBa_B0032

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 target gene CheZ.

2.Then link the skeleton to target gene CheZ-TT.

3.Next, link RBS-CheZ-TT with promoter Plac.

Characterization stage

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


Reference