Difference between revisions of "Part:BBa K395302:Experience"
Line 11: Line 11:
 
|-
 
|-
 
|width='10%'|
 
|width='10%'|
−
<partinfo>BBa_K395302 AddReview number</partinfo>
+
<I>Tokyo Tech iGEM2010</I>
−
<I>Username</I>
+
 
|width='60%' valign='top'|
 
|width='60%' valign='top'|
−
Enter the review inofrmation here.
+
[[Image:Tokyotech_ompc_graph.jpg|thumb|center|400px|Figure 1. Induction of new'' OmpC'' series in high osmolarity medium  Tokyo Tech iGEM2010]]
 +
In order to characterize P''ompC(C)'' [https://parts.igem.org/Part:BBa_K395301 BBa_395301], P''ompC(CB)'' [https://parts.igem.org/Part:BBa_K395302 BBa395302] and P''ompC(CS1)'' [https://parts.igem.org/Part:BBa_K395303 BBa_395303], each promoter was attached to GFP and its transcriptional activity was measured through the GFP expression. <br><br>
 +
P''ompC(C)''-GFP[https://parts.igem.org/Part:BBa_K395304 BBa_395304] , P''ompC(CB)''-GFP [https://parts.igem.org/Part:BBa_K395305 BBa_395305], P''ompC(CS1)''-GFP [https://parts.igem.org/Part:BBa_K395306 BBa_395306]and P''ompC(WT)''-GFP[https://parts.igem.org/Part:BBa_K395307 BBa_395307] on pSB3K3 was introduced into E. coli strain MG1655. A strain containing placI<sup>q</sup>-GFP, plasmid (BBa_J54202), a constitutive GFP expressive promoter and promoterless GFP reporter plasmid (BBa_J54103) were used as a positive control and negative control respectively. <br><br>
 +
Overnight cultures of reporter strains grown at 37 °C in Medium A containing appropriated antibiotics were diluted at least 1:100 in the medium and incubated at 37 °C as fresh cultures. After their OD<sub>590</sub> reached 0.2, the fresh culture was diluted 1 : 3 into 2 ml of pre-warmed medium A. For high osmolarity conditions, the cultures were diluted with sucrose supplemented medium to the final concentration of 15% (wt/vol). After 4 hours of induction, fluorescence intensity was measured with fluorometer. <br><br>
 +
After 4 hours of high osmolarity induction by sucrose, transcriptional activity of P''ompC(CB)''-GFP and P''ompC(CS1)''-GFP increased 2.5 folds and 2.3 folds respectively. However, significant amount of leaky expression was found in P''ompC(CS1)''-GFP without induction. In contrast, under the same conditions, we found no significant difference of GFP expression in P''ompC(C)''-GFP and P''ompC(WT)''-GFP. [http://2010.igem.org/Team:Tokyo_Tech/Project/wolf_coli/New_Series_of_PompC ...see more about P''ompC'' series]
 +
 
 +
|};
 +
<!-- End of the user review template -->the review inofrmation here.
 
|};
 
|};
 
<!-- End of the user review template -->
 
<!-- End of the user review template -->
 
<!-- DON'T DELETE --><partinfo>BBa_K395302 EndReviews</partinfo>
 
<!-- DON'T DELETE --><partinfo>BBa_K395302 EndReviews</partinfo>

Revision as of 13:58, 26 October 2010

Characterization of new series of OmpC propmoters
Figure 1. The induction of new OmpC series in high osmolarity medium

Applications of BBa_K395302

User Reviews

UNIQ2125a2c12b495e54-partinfo-00000000-QINU the review inofrmation here. |}; UNIQ2125a2c12b495e54-partinfo-00000001-QINU