Difference between revisions of "Part:BBa K1412801"
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BBa_K1412801: Plac-RBS(0.01)-<i>CheZ</i>-TT
 
BBa_K1412801: Plac-RBS(0.01)-<i>CheZ</i>-TT
  
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This part consists of a <i>[http://en.wikipedia.org/wiki/Chemotaxis CheZ]</i><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 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.
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This part consists of a <i>[http://en.wikipedia.org/wiki/Chemotaxis 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 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.
  
  
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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(Plac-RBS(changeable)-<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>.
 
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(Plac-RBS(changeable)-<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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==='''Relevant parts'''===
 
==='''Relevant parts'''===
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----
 
----
  
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==='''Source'''===
  
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===Source===
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<bbpart>BBa_R0010</bbpart>
  
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<bbpart>BBa_B0033</bbpart>
  
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----
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<bbpart>BBa_K629003</bbpart>
  
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<bbpart>BBa_R0010</bbpart> <bbpart>BBa_B0033</bbpart> <bbpart>BBa_K629003</bbpart> <bbpart>BBa_B0015</bbpart>
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<bbpart>BBa_B0015</bbpart>
  
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here
 
===Usage and Biology===
 
===Usage and Biology===
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<!-- -->
 
<!-- -->
 
<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K1412801 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K1412801 SequenceAndFeatures</partinfo>
  
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===Protocol===
 
  
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==='''Protocol'''===
  
 
----
 
----
  
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Genetic link stage
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===='''Genetic link stage'''====
  
 
1.As a skeleton, TT terminator link with target gene <i>CheZ</i>.
 
1.As a skeleton, TT terminator link with target gene <i>CheZ</i>.
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3.Next, link  RBS-<i>CheZ</i>-TT with promoter Plac.
 
3.Next, link  RBS-<i>CheZ</i>-TT with promoter Plac.
  
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===='''Characterization stage'''====
  
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Characterization stage
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1.Transfer the part Plac-RBS-<i>CheZ</i>-TT into <i>E.coli</i> (<i>CheZ</i> knocked out), and coat plates, culture for hours to measure the migration diameter of <i>E.coli</i>.
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1.Transfer the part Plac-RBS-<i>CheZ</i>-TT into <i>E.coli</i> (<i>CheZ</i> knocked out), and coat plates, culture for hours to measure  
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the migration diameter of <i>E.coli</i>.
 
  
 
==='''Reference'''===
 
==='''Reference'''===
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----
 
----
  
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<!-- Uncomment this to enable Functional Par
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<!-- Uncomment this to enable Functional Parameter display  
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ameter display  
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===Functional Parameters===
 
===Functional Parameters===
 
<partinfo>BBa_K1412801 parameters</partinfo>
 
<partinfo>BBa_K1412801 parameters</partinfo>
 
<!-- -->
 
<!-- -->

Revision as of 08:29, 6 October 2014

Characterize efficiency of RBS with chemotaxis

BBa_K1412801: Plac-RBS(0.01)-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, 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 knocked 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_K1412014

BBa_K1412614

BBa_K1412829


Notes

Source

BBa_R0010

BBa_B0033

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