Difference between revisions of "Part:BBa K1412014"

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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K1412014 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K1412014 SequenceAndFeatures</partinfo>
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==='''Protocol'''===
 
==='''Protocol'''===

Revision as of 18:33, 16 October 2014

Characterize the efficiency of pTetR (R0040) with chemotaxis

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

This part consists of [http://en.wikipedia.org/wiki/Chemotaxis CheZ] gene which can express CheZ protein make E.coli tumble or swim straight. In this light, we can characterize the efficiency of promoter by replacing 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 connect the promoter with GFP, then measuring the fluorescence intensity of GFP. In our part, you just need connect RBS(1.0) after a pTetR promoter and before CheZ gene, ending with double 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


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

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

1. Kelly J R, Rubin A J, Davis J H, et al. Measuring the activity of BioBrick promoters using an in vivo reference standard[J]. Journal of biological engineering, 2009, 3(1): 4.

2. https://parts.igem.org/Part:BBa_R0010:Experience


More information, click here: [http://2014.igem.org/Team:XMU-China# XMU-China]