Difference between revisions of "Part:BBa K1412801"
(→‎Protocol)
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Genetic link stage
 
Genetic link stage
  
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1.As a skeleton, <i>TT</i> terminator link with <i>CheZ</i>.
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1.As a skeleton, TT terminator link with target gene <i>CheZ</i>.
  
−
2.Then the skeleton <i>CheZ-TT</i> link to <i>RBS</i>.
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2.Then link the skeleton to target gene <i>CheZ</i>-TT.
 +
 
 +
3.Next, link RBS-<i>CheZ</i>-TT with promoter Plac.
  
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3.Next, link  <i>RBS-CheZ-TT</i> with promoter <i>Plac</i>.
 
  
 
Characterization stage
 
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  
+
1.Transfer the part Plac-RBS-CheZ-TT into E.coli (CheZ knocked out), and coat plates, culture for hours to measure the migration
  
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migration diameter of E.coli.
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diameter of E.coli.
  
 
==='''Reference'''===
 
==='''Reference'''===

Revision as of 18:04, 3 October 2014

Characterize efficiency of RBS with chemotaxis

BBa_K1412801: Plac-RBS(0.01)-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 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.


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_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