Difference between revisions of "Part:BBa K1412829"
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BBa_K1412829: <i>Plac-RBS(0.01)-CheZ-TT</i> | BBa_K1412829: <i>Plac-RBS(0.01)-CheZ-TT</i> | ||
| − | 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 | + | 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 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 CL-1</i> (<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 CL-1</i> (<i>CheZ</i> knocked out), and coat plates, culture on semi-solid medium to measure the migration diameter of <i>E.coli</i>. |
==='''Relevant parts'''=== | ==='''Relevant parts'''=== | ||
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genetic link stage | genetic link stage | ||
| − | 1.As a skeleton, | + | 1.As a skeleton, TT terminator link with <i>CheZ</i>. |
| − | 2.Then the skeleton <i>CheZ | + | 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 E.coli (<i>CheZ</i> knocked out), and coat plates, culture for hours to measure the migration diameter of E.coli. |
Revision as of 04:45, 3 October 2014
Characterize efficiency of RBS with chemotaxis
BBa_K1412829: 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.
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 3H:13-P3-3H BBa_R0010 2L:14-P2-2L BBa_B0033 18G:14-P1-18G BBa_K629003 4F:13-P3-4F BBa_B0015
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 knocked out), and coat plates, culture for hours to measure the migration diameter of E.coli.