Difference between revisions of "Part:BBa K1412614"

Revision as of 08:48, 6 October 2014

Characterize the efficiency of promoter(BBa_K206000) with chemotaxis

BBa_K1412614: pBAD-RBS(1)-CheZ-TT

This part consists of a [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 efficiency of promoter by just change different promoters before a CheZ gene. Then we can characterize the efficiency of promoter via measuring the migration distance positively associated with the expression strength of CheZ protein.

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 knocked out), and coat plates, culuture on semi-solid medium to measure the migration diameter of E.coli.

Relevant parts

Notes

Source

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 125
21
INCOMPATIBLE WITH RFC[21]
Illegal BamHI site found at 65
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

Protocol

Genetic link stage

1.As a skeleton, TT terminator link with CheZ.

2.Then the skeleton CheZ-TT link to RBS.

3.Next, link RBS-CheZ-TT with promoter Plac.

Characterization stage

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

@@ Line 6: / Line 6: @@
 '''BBa_K1412614: pBAD-RBS(1)-<i>CheZ</i>-TT'''
-This part consists of a <i>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 before a <i>CheZ</i> gene. Then we can characterize the efficiency of promoter via measuring the migration distance positively associated with the expression strength of CheZ protein.
+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 before a <i>CheZ</i> gene. Then we can characterize the efficiency of promoter via measuring the migration distance positively associated with the expression strength of CheZ protein.
 [[File:Characterization_process_design.png |500px]]
 ==='''Usage'''===
@@ Line 53: / Line 54: @@
 <!-- Add more about the biology of this part here
 ===Usage and Biology===
-----
 <!-- -->
 <span class='h3bb'>Sequence and Features</span>
 <partinfo>BBa_K1412614 SequenceAndFeatures</partinfo>
-===='''Protocol'''====
-====Genetic link stage====
+==='''Protocol'''===
+----
+===='''Genetic link stage'''====
 .As a skeleton, TT terminator link with <i>CheZ</i>.
@@ Line 70: / Line 71: @@
 .Next, link  RBS-<i>CheZ</i>-TT with promoter Plac.
-====Characterization stage====
+===='''Characterization stage'''====
-.Transfer the part Plac-RBS-<i>CheZ</i>-TT into <i>E.coli</i> (<i>CheZ</i> knock out), and coat plates, culture for hours to measure the
+.Transfer the part Plac-RBS-<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>.
-migration diameter of <i>E.coli</i>.
 ==='''Reference'''===