Difference between revisions of "Part:BBa K1412829"

Revision as of 20:58, 17 October 2014

Characterize efficiency of RBS with chemotaxis

BBa_K1412829: Plac-RBS(0.3)-cheZ-TT

This part consists of [http://en.wikipedia.org/wiki/Chemotaxis cheZ] gene which can express CheZ protein makes E.coli tumble or swim straight. In this light, we can characterize the RBS efficiency by just change different RBS. Then we can characterize the efficiency of RBS via measuring the migration distance positively associated with the expression strength of CheZ.

Figure 1.The schematic diagram of how we characterize the activity of promoters.

Usage

When we want to characterize the efficiency of RBS, we usually connect the RBS with GFP, and then measure the fluorescence intensity of GFP. In our part, you only need to connect RBS after a Plac promoter and before cheZ gene, ending with a double terminator(Plac-RBS(changeable)-cheZ-TT). Then transferred this gene circuit into E.coli CL-1 (cheZ knocked out), and coated plates, cultured on semi-solid medium to measure the migration diameter of E.coli.

Relevant parts

BBa_K1412000: pLac-RBS(1.0)-cheZ-TT cheZ generator under pLac promoter

BBa_K1412005: RBS(1.0)-cheZ-TT

BBa_K1412006: RBS(0.01)-cheZ-TT

BBa_K1412007: RBS(0.3)-cheZ-TT

BBa_K1412014: pTet-RBS(1.0)-cheZ-TT Characterize the efficiency of promoters with chemotaxis

BBa_K1412614: pBAD-RBS(1.0)-cheZ-TT Characterize the efficiency of promoters with chemotaxis

BBa_K1412801: pLac-RBS(0.01)-cheZ-TT Characterize the efficiency of RBS with chemotaxis

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

Reference

More information, click here: [http://2014.igem.org/Team:XMU-China# XMU-China]

@@ Line 6: / Line 6: @@
 '''BBa_K1412829: Plac-RBS(0.3)-<i>cheZ</i>-TT'''
-This part consists of <i>[http://en.wikipedia.org/wiki/Chemotaxis cheZ]</i> gene which can express CheZ protein make <i>E.coli</i> 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 <i>[http://en.wikipedia.org/wiki/Chemotaxis cheZ]</i> gene which can express CheZ protein makes <i>E.coli</i> tumble or swim straight. In this light, we can characterize the RBS efficiency by just change different RBS. Then we can characterize the efficiency of RBS via measuring the migration distance positively associated with the expression strength of CheZ.
-[[File:]]
 [[File:Characterization_process_design.png |700px|thumb|center|<b>Figure 1</b>.The schematic diagram of how we characterize the activity of promoters.]]
@@ Line 17: / Line 16: @@
 ----
-When we want to characterize the efficiency of RBS, we usually connect the RBS with GFP, and then measure the fluorescence intensity of GFP. In our part, you only need to connect RBS after a Plac promoter and before  <i>cheZ</i> gene, ending with a double 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 connect the RBS with GFP, and then measure the fluorescence intensity of GFP. In our part, you only need to connect RBS after a Plac promoter and before  <i>cheZ</i> gene, ending with a double terminator(Plac-RBS(changeable)-<i>cheZ</i>-TT). Then transferred this gene circuit into <i>E.coli</i> CL-1 (<i>cheZ</i> knocked out), and coated plates, cultured on semi-solid medium to measure the migration diameter of <i>E.coli</i>.