Difference between revisions of "Part:BBa K1412801"

Latest revision as of 00:30, 18 October 2014

Characterize efficiency of RBS with chemotaxis

BBa_K1412801: pLac-RBS(0.01)-cheZ-TT

This part consists of [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 replacing 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.

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

Usage

When we want to characterize the efficiency of RBS, we usually connect the RBS with GFP, then measuring the fluorescence intensity of GFP. In our part, you need just connect RBS after a pLac promoter and before a cheZ gene, ending with double terminator(pLac-RBS(changeable)-cheZ-TT). Then transferred this gene circuit into E. coli (cheZ knocked out), and coated plates, culutured 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_K1412829: Plac-RBS(0.3)-cheZ-TT Characterize 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 4: / Line 4: @@
 ----
-BBa_K1412801: Plac-RBS(0.01)-<i>CheZ</i>-TT
+'''BBa_K1412801: pLac-RBS(0.01)-<i>cheZ</i>-TT'''
-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.
+This part consists of <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 replacing 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.
-[[File:Characterization_process_design.png |500px]]
+[[File:Characterization.jpg |800px|thumb|center|<b>Figure 1</b>. The schematic diagram of how we characterize the activity of RBS]]
@@ Line 16: / Line 16: @@
 ----
-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>.
+When we want to characterize the efficiency of RBS, we usually connect the RBS with GFP, then measuring the fluorescence intensity of GFP. In our part, you need just connect [https://parts.igem.org/wiki/index.php/Part:BBa_B0032 RBS] after a [https://parts.igem.org/wiki/index.php/Part:BBa_R0010 pLac] promoter and before a <i>[https://parts.igem.org/Part:BBa_K629003 cheZ]</i> gene, ending with double terminator(pLac-RBS(changeable)-<i>cheZ</i>-TT). Then transferred this gene circuit into <i>E. coli</i> (<i>cheZ</i> knocked out), and coated plates, culutured on semi-solid medium to measure the migration diameter of <i>E. coli</i>.
 ==='''Relevant parts'''===
@@ Line 23: / Line 22: @@
 ----
-<bbpart>BBa_K1412000</bbpart>
+<bbpart>BBa_K1412000</bbpart>: pLac-RBS(1.0)-<i>cheZ</i>-TT CheZ generator under pLac promoter
-<bbpart>BBa_K1412005</bbpart>
+<bbpart>BBa_K1412005</bbpart>: RBS(1.0)-<i>cheZ</i>-TT
-<bbpart>BBa_K1412006</bbpart>
+<bbpart>BBa_K1412006</bbpart>: RBS(0.01)-<i>cheZ</i>-TT
-<bbpart>BBa_K1412007</bbpart>
+<bbpart>BBa_K1412007</bbpart>: RBS(0.3)-<i>cheZ</i>-TT
-<bbpart>BBa_K1412014</bbpart>
+<bbpart>BBa_K1412014</bbpart>: pTet-RBS(1.0)-cheZ-TT Characterize the efficiency of promoters with chemotaxis
-<bbpart>BBa_K1412614</bbpart>
+<bbpart>BBa_K1412614</bbpart>: pBAD-RBS(1.0)-cheZ-TT Characterize the efficiency of promoters with chemotaxis
-<bbpart>BBa_K1412829</bbpart>
+<bbpart>BBa_K1412829</bbpart>: Plac-RBS(0.3)-cheZ-TT Characterize efficiency of RBS with chemotaxis
-==='''Notes'''===
-----
-==='''Source'''===
-<bbpart>BBa_R0010</bbpart>
-<bbpart>BBa_B0033</bbpart>
-<bbpart>BBa_K629003</bbpart>
-<bbpart>BBa_B0015</bbpart>
@@ Line 60: / Line 44: @@
-==='''Protocol'''===
+==='''Reference'''===
 ----
-===='''Genetic link stage'''====
+<I><B>More information, click here: [http://2014.igem.org/Team:XMU-China# XMU-China]
-.As a skeleton, TT terminator link with target gene <i>CheZ</i>.
-.Then link the skeleton to target gene <i>CheZ</i>-TT.
-.Next, link  RBS-<i>CheZ</i>-TT with promoter Plac.
-===='''Characterization stage'''====
-.Transfer the part Plac-RBS-<i>CheZ</i>-TT into <i>E.coli</i> (<i>CheZ</i> knocked out), and coat plates, culture for hours to measure the migration diameter of <i>E.coli</i>.
-==='''Reference'''===
-----
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