Difference between revisions of "Part:BBa K1412014"
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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 <i>CheZ</i> gene, ending with double terminator(pTetR-RBS(1.0)-<i>CheZ</i>-TT). Then transfer this gene circuit into <i>E.coli</i> (<i>CheZ</i> knocked out), and coat plates, culuture on semi-solid medium to measure the migration diameter of <i>E.coli</i>.
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When we want to characterize the activity of promoter which have not been measured, we usually connect the promoter with GFP, then measuring the fluorescence intensity of GFP. In our project, We linked those three parts: <bbpart>BBa_K1412000</bbpart>, <bbpart>BBa_K1412014</bbpart> and <bbpart>BBa_K1412614]</bbpart>. Then we transferred those gene circuits into <i>E. coli</i> (<i>cheZ</i> knocked out), coated plates, and cultured on semi-solid medium to measure the migration diameter of <i>E. coli</i> parallelly. We verified whether the activity of pLac and pTetR we measured were consistent with which had been measured[1][2] in the beginning. If so, it means that our method is the feasible and we can use this method to characterize the activity of pBAD.
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==='''Relevant parts'''===
 
==='''Relevant parts'''===

Revision as of 19:18, 17 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 tumbling or swimming straight. In this light, we linked different promoters before cheZ gene. Then we can characterize the efficiency of promoter via measuring the migration distance positively associated with the expression strength of CheZ protein.


Figure 1. The schematic diagram of how we characterize the activity of promoters.


Usage

When we want to characterize the activity of promoter which have not been measured, we usually connect the promoter with GFP, then measuring the fluorescence intensity of GFP. In our project, We linked those three parts: BBa_K1412000, BBa_K1412014 and BBa_K1412614. Then we transferred those gene circuits into E. coli (cheZ knocked out), coated plates, and cultured on semi-solid medium to measure the migration diameter of E. coli parallelly. We verified whether the activity of pLac and pTetR we measured were consistent with which had been measured[1][2] in the beginning. If so, it means that our method is the feasible and we can use this method to characterize the activity of pBAD.

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]

Reference

[1] [http://www.jbioleng.org/content/3/1/4 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]