Difference between revisions of "Part:BBa K1412014"

Revision as of 17:40, 14 October 2014

Characterize the efficiency of pTetR (R0040) with chemotaxis

BBa_K1412014: pTetR-RBS(1.0)-CheZ-TT

This part is consist of [http://en.wikipedia.org/wiki/Chemotaxis CheZ] gene which can express CheZ protein make E.coli tumble or swim straight. In this light, we can characterize the efficiency of promoter by replacing 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 promoter, we usually connect the promoter with GFP, then measuring the fluorescence intensity of GFP. In our part, you just need connect RBS(1.0) after a pTetR promoter and before CheZ gene, ending with double terminator(pTetR-RBS(1.0)-CheZ-TT). Then transfer this gene circuit into E.coli (CheZ knocked out), and coat plates, culuture 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_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

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]

Protocol

Genetic connection stage

1.As a backbone, double terminator connect with CheZ.

2.Then the backbone CheZ-TT connect with RBS(1.0).

3.Next, connect RBS(1.0)-CheZ-TT with promoter pTetR.

Characterization stage

1. Transfer the part pTetR-RBS(1.0)-CheZ-TT into E.coli (CheZ knock out), and coat plates, culture for hours to measure the migration diameter of E.coli.

Reference

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

@@ Line 4: / Line 4: @@
 ----
-BBa_K1412014: pTetR-RBS(1.0)-<i>CheZ</i>-TT
+'''BBa_K1412014: pTetR-RBS(1.0)-<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 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.
+This part is consist 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 efficiency of promoter by replacing different promoters. Then we can characterize the efficiency of promoter via measuring the migration distance positively associated with the expression strength of CheZ.
@@ Line 16: / Line 16: @@
 ----
-When we want to characterize the efficiency of promoter, we usually link the promoter with GFP, then characterize the promoter by just measure the fluorescence intensity of GFP. In our part, you need just link RBS(1.0) after a pTetR promoter and before a <i>CheZ</i> gene, ending with a TT terminator(pTetR-RBS(1.0)-<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 promoter, we usually connect the promoter with GFP, then measuring the fluorescence intensity of GFP. In our part, you just need connect RBS(1.0) after a pTetR promoter and before <i>CheZ</i> gene, ending with double terminator(pTetR-RBS(1.0)-<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>.
@@ Line 36: / Line 36: @@
 <bbpart>BBa_K1412829</bbpart>: pLac-RBS(0.3)-CheZ-TT Characterize efficiency of RBS with chemotaxis
-==='''Notes'''===
-----
-===='''Source'''====
-<bbpart>BBa_R0040</bbpart>
-<bbpart>BBa_B0034</bbpart>
-<bbpart>BBa_K629003</bbpart>
-<bbpart>BBa_B0015</bbpart>
@@ Line 65: / Line 49: @@
 ----
-===='''Genetic link stage'''====
+===='''Genetic connection stage'''====
-.As a skeleton, TT terminator link with <i>CheZ</i>.
+.As a backbone, double terminator connect with <i>CheZ</i>.
-.Then the skeleton <i>CheZ</i>-TT link to RBS(1.0).
+.Then the backbone <i>CheZ</i>-TT connect with RBS(1.0).
-.Next, link  RBS(1.0)-<i>CheZ</i>-TT with promoter pTetR.
+.Next, connect RBS(1.0)-<i>CheZ</i>-TT with promoter pTetR.
 ===='''Characterization stage'''====
-.Transfer the part pTetR-RBS(1.0)-<i>CheZ</i>-TT into <i>E.coli</i> (<i>CheZ</i> knock out), and coat plates, culture for hours to measure the migration diameter of <i>E.coli</i>.
+. Transfer the part pTetR-RBS(1.0)-<i>CheZ</i>-TT into <i>E.coli</i> (<i>CheZ</i> knock out), and coat plates, culture for hours to measure the migration diameter of <i>E.coli</i>.
@@ Line 81: / Line 65: @@
 ----
+<I><B>More information, click here: [http://2014.igem.org/Team:XMU-China# XMU-China]
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