Difference between revisions of "Part:BBa K1412014"
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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 E.coli (<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>. |
==='''Relevant parts'''=== | ==='''Relevant parts'''=== | ||
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2.Then the skeleton <i>CheZ</i>-TT link to RBS. | 2.Then the skeleton <i>CheZ</i>-TT link to RBS. | ||
| − | 3.Next, link <i> | + | 3.Next, link RBS-<i>CheZ</i>-TT with promoter Plac. |
'''Characterization stage''' | '''Characterization stage''' | ||
| − | 1.Transfer the part | + | 1.Transfer the part Plac-RBS-<i>CheZ</i>-TT into <i>E.coli</i> (<i>CheZ</i> knock out), and coat plates, culture for hours to measure the migration diameter of <i>E.coli</i>. |
| − | + | ||
| − | migration diameter of E.coli. | + | |
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| − | <!-- Uncomment this to enable Functional | + | <!-- Uncomment this to enable Functional Parameter display |
| − | + | ||
===Functional Parameters=== | ===Functional Parameters=== | ||
<partinfo>BBa_K1412014 parameters</partinfo> | <partinfo>BBa_K1412014 parameters</partinfo> | ||
<!-- --> | <!-- --> | ||
Revision as of 08:10, 6 October 2014
Characterize the efficiency of pTetR (R0040) with chemotaxis
BBa_K1412014: pTet-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 promoter by 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 knocked out), and coat plates, culuture 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 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