Difference between revisions of "Part:BBa K1412014"

 
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<partinfo>BBa_K1412014 short</partinfo>
 
<partinfo>BBa_K1412014 short</partinfo>
  
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This part consists of a CheZ gene which can express CheZ protein deciding E.coli whether tumble or swim straight.In this light, we can characterize the efficiency of promoters by just change different promoters.Then we can characterize the efficiency of RBS via measuring the migration distance positively associated with the expression strength of CheZ.
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BBa_K1412801: <i>pTETR-RBS(1)-CheZ-TT</i>
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This part consists of a <i>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 promoterby just change different promoters.Then we can characterize the efficiency of promoter via measuring the migration distance positively associated with the expression strength of <i>CheZ</i>.
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==='''Usage'''===
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When we want to characterize the efficiency of RBS, we usually link the RBS with GFP, then characterize the RBS by just measure the fluorescence intensity of GFP. In our part, you need just link RBS after a Plac promoter and before a <i>CheZ </i> gene, ending with a TT terminator(<i>Plac-RBS</i>(changeable)-<i>CheZ-TT</i>). Then transfer this gene circuit into E.coli (<i>CheZ</i> knock out), and coat plates, culuture on semi-solid medium to measure the migration diameter of <i>E.coli</i>.
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==='''Relevant parts'''===
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<bbpart>BBa_K1412000</bbpart>
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<bbpart>BBa_K1412005</bbpart>
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<bbpart>BBa_K1412006</bbpart>
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<bbpart>BBa_K1412007</bbpart>
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<bbpart>BBa_K1412614</bbpart>
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<bbpart>BBa_K1412801</bbpart>
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<bbpart>BBa_K1412829</bbpart>
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==='''Notes'''===
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Source
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6F:14-P2-6F <bbpart>BBa_R0040</bbpart>
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2M:13-P5-2M <bbpart>BBa_B0034</bbpart>
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18G:14-P1-18G <bbpart>BBa_K629003</bbpart>
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4F:13-P3-4F <bbpart>BBa_B0015</bbpart>
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K1412014 SequenceAndFeatures</partinfo>
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<partinfo>BBa_K1412829 SequenceAndFeatures</partinfo>
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===Protocol===
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genetic link stage
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1.As a skeleton, <i>TT</i> terminator link with <i>CheZ</i>.
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2.Then the skeleton <i>CheZ-TT</i> link to <i>RBS</i>.
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3.Next, link  <i>RBS-CheZ-TT</i> with promoter <i>Plac</i>.
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characterization stage
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1.Transfer the part <i>Plac-RBS-CheZ-TT</i> into E.coli (<i>CheZ</i> knock out), and coat plates, culture for hours to measure the
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migration diameter of E.coli.
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==='''Reference'''===
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ameter display  
 
===Functional Parameters===
 
===Functional Parameters===
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<partinfo>BBa_K1412014 parameters</partinfo>
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<partinfo>BBa_K1412801 parameters</partinfo>
 
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Revision as of 03:48, 2 October 2014

Characterize the efficiency of pTetR (R0040) with chemotaxis

BBa_K1412801: pTETR-RBS(1)-CheZ-TT

This part consists of a CheZ gene which can express CheZ protein deciding E.coli whether tumble or swim straight.In this light, we can characterize the efficiency of promoterby 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.


Usage

When we want to characterize the efficiency of RBS, we usually link the RBS with GFP, then characterize the RBS by just measure the fluorescence intensity of GFP. In our part, you need just 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 (CheZ knock out), and coat plates, culuture on semi-solid medium to measure the migration diameter of E.coli.


Relevant parts

BBa_K1412000

BBa_K1412005

BBa_K1412006

BBa_K1412007

BBa_K1412614

BBa_K1412801

BBa_K1412829


Notes

Source 6F:14-P2-6F BBa_R0040 2M:13-P5-2M BBa_B0034 18G:14-P1-18G BBa_K629003 4F:13-P3-4F 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.

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

characterization stage

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

migration diameter of E.coli.


Reference