Difference between revisions of "Part:BBa K1412829"

Revision as of 06:23, 1 October 2014

Characterize efficiency of RBS with chemotaxis

BBa_K1412829: 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

Notes

Source 3H:13-P3-3H BBa_R0010

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 genetic link stage
+.As a skeleton, <i>TT</i> terminator link with <i>CheZ</i>.
+.Then the skeleton <i>CheZ-TT</i> link to <i>RBS</i>.
+.Next, link  <i>RBS-CheZ-TT</i> with promoter <i>Plac</i>.
 characterization stage
+.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 migration diameter of E.coli.