Difference between revisions of "Part:BBa K1412829"

Revision as of 17:22, 14 October 2014

Characterize efficiency of RBS with chemotaxis

BBa_K1412829: Plac-RBS(0.3)-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 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 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 transfer this gene circuit into E.coli CL-1 (CheZ knocked out), and coat plates, culture 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]

Protocol

Genetic connection stage

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

2.Then connect the backbone to target gene CheZ-TT.

3.Next, connect RBS-CheZ-TT with promoter pLac.

Characterization stage

1.Transfer the part Plac-RBS-CheZ-TT into E.coli (CheZ knocked 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_K1412829: Plac-RBS(0.3)-<i>CheZ</i>-TT
+'''BBa_K1412829: Plac-RBS(0.3)-<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 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 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.
@@ Line 16: / Line 16: @@
 ----
-When we want to characterize the efficiency of RBS, we usually link the RBS with GFP, and then characterize the RBS by just measure the fluorescence intensity of GFP. In our part, you only need to 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> 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 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>.
@@ Line 36: / Line 36: @@
 <bbpart>BBa_K1412801</bbpart>: pLac-RBS(0.01)-CheZ-TT Characterize the efficiency of RBS with chemotaxis
-==='''Notes'''===
-----
-===='''Source'''====
-<bbpart>BBa_R0010</bbpart>
-<bbpart>BBa_B0032</bbpart>
-<bbpart>BBa_K629003</bbpart>
-<bbpart>BBa_B0015</bbpart>
@@ Line 64: / Line 49: @@
 ----
-===='''Genetic link stage'''====
+===='''Genetic connection stage'''====
-.As a skeleton, TT terminator link with target gene <i>CheZ</i>.
+.As a backbone, double terminator connect with target gene <i>CheZ</i>.
-.Then link the skeleton to target gene <i>CheZ</i>-TT.
+.Then connect the backbone to target gene <i>CheZ</i>-TT.
-.Next, link  RBS-<i>CheZ</i>-TT with promoter pLac.
+.Next, connect RBS-<i>CheZ</i>-TT with promoter pLac.
@@ Line 83: / Line 68: @@
 ----
+<I><B>More information, click here: [http://2014.igem.org/Team:XMU-China# XMU-China]
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