Difference between revisions of "Part:BBa K1412014"
| Line 2: | Line 2: | ||
<partinfo>BBa_K1412014 short</partinfo> | <partinfo>BBa_K1412014 short</partinfo> | ||
| − | BBa_K1412801: | + | BBa_K1412801: pTETR-RBS(1)-<i>CheZ</i>-TT |
| − | This part consists of a <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 efficiency of | + | This part consists of a <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 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 <i>CheZ</i>. |
| Line 33: | Line 33: | ||
==='''Notes'''=== | ==='''Notes'''=== | ||
| − | + | ====Source==== | |
| − | Source | + | |
14-P2-6F <bbpart>BBa_R0040</bbpart> 13-P5-2M <bbpart>BBa_B0034</bbpart> 14-P1-18G <bbpart>BBa_K629003</bbpart> 13-P3-4F <bbpart>BBa_B0015</bbpart> | 14-P2-6F <bbpart>BBa_R0040</bbpart> 13-P5-2M <bbpart>BBa_B0034</bbpart> 14-P1-18G <bbpart>BBa_K629003</bbpart> 13-P3-4F <bbpart>BBa_B0015</bbpart> | ||
| Line 47: | Line 46: | ||
===Protocol=== | ===Protocol=== | ||
| − | + | ====Genetic link stage==== | |
1.As a skeleton, <i>TT</i> terminator link with <i>CheZ</i>. | 1.As a skeleton, <i>TT</i> terminator link with <i>CheZ</i>. | ||
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3.Next, link <i>RBS-CheZ-TT</i> with promoter <i>Plac</i>. | 3.Next, link <i>RBS-CheZ-TT</i> with promoter <i>Plac</i>. | ||
| − | + | ====Characterization stage==== | |
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 <i>Plac-RBS-CheZ-TT</i> into E.coli (<i>CheZ</i> knock out), and coat plates, culture for hours to measure the | ||
Revision as of 15:14, 3 October 2014
Characterize the efficiency of pTetR (R0040) with chemotaxis
BBa_K1412801: pTETR-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 knock out), and coat plates, culuture on semi-solid medium to measure the migration diameter of E.coli.
Relevant parts
Notes
====Source====
14-P2-6F BBa_R0040 13-P5-2M BBa_B0034 14-P1-18G BBa_K629003 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 knock out), and coat plates, culture for hours to measure the
migration diameter of E.coli.