Composite

Part:BBa_K1412614

Designed by: Ruihua Zhang   Group: iGEM14_XMU-China   (2014-10-01)
Revision as of 07:57, 17 October 2014 by D (Talk | contribs) (→‎Usage)

Characterize the efficiency of promoter(BBa_K206000) with chemotaxis

BBa_K1412614: pBAD- 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 can characterize the efficiency of promoter by replacing 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.


Characterization process design.png


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 link this third parts: BBa_K1412000, BBa_K1412014 and BBa_K1412614. Then we transfer those gene circuits into E.coli (CheZ knocked out), coat plates, and culture on semi-solid medium to measure the migration diameter of E.coli parallelly. Firstly, we verify whether the activity of pLac and pTet we measured were consistent with which had been measured[1][2]. 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_K1412014: pTetR-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
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 125
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 65
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

Reference

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

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