Difference between revisions of "Part:BBa K1412829"
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<partinfo>BBa_K1412829 short</partinfo> | <partinfo>BBa_K1412829 short</partinfo> | ||
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| − | + | BBa_K1412829: Plac-RBS(0.3)-<i>CheZ</i>-TT | |
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. | 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. | ||
| + | [[File:Characterization_process_design.png |500px]] | ||
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| − | When we want to characterize the efficiency of RBS, we usually link the RBS with GFP, and then characterize the RBS by just measure the fluorescence intensity of GFP. In our part, you only need to 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> CL-1 (<i>CheZ</i> knocked out), and coat plates, culture 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, and then characterize the RBS by just measure the fluorescence intensity of GFP. In our part, you only need to 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> CL-1 (<i>CheZ</i> knocked out), and coat plates, culture 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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<!-- 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_K1412829 SequenceAndFeatures</partinfo> | <partinfo>BBa_K1412829 SequenceAndFeatures</partinfo> | ||
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==='''Protocol'''=== | ==='''Protocol'''=== | ||
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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. | ||
| − | ====''' | + | ===='''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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==='''Reference'''=== | ==='''Reference'''=== | ||
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<!-- Uncomment this to enable Functional Par | <!-- Uncomment this to enable Functional Par | ||
Revision as of 08:30, 6 October 2014
Characterize efficiency of RBS with chemotaxis
BBa_K1412829: Plac-RBS(0.3)-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.
Usage
When we want to characterize the efficiency of RBS, we usually link the RBS with GFP, and then characterize the RBS by just measure the fluorescence intensity of GFP. In our part, you only need to 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 CL-1 (CheZ knocked out), and coat plates, culture on semi-solid medium to measure the migration diameter of E.coli.
Relevant parts
Notes
Source
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE 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