Difference between revisions of "Part:BBa K395301:Experience"

Revision as of 12:04, 26 October 2010

This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

Applications of BBa_K395301

User Reviews

UNIQ1bf67392673b4b26-partinfo-00000000-QINU

Tokyo Tech iGEM2010

Figure 1. Induction of new OmpC series in high osmolarity medium

PompC(C)-GFP, PompC(CB)-GFP, PompC(CS1)-GFP and PompC(WT)-GFP on pSB3K3 was introduced into E. coli strain MG1655. A strain containing placI^q-GFP, plasmid (BBa_J54202), a constitutive GFP expressive promoter, and promoterless GFP reporter plasmid (BBa_J54103) was used as a positive control and negative control respectively.

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 OD590 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.

After 4 hours of sucrose induction, transcriptional activity of PompC(CB)-GFP and PompC(CS1)-GFP increased 2.5 folds and 2.3 folds respectively. However, significant amount of leaky expression was found in PompC(CS1)-GFP without induction. In contrast, under the same conditions, we found no significant difference of GFP expression in PompC(C)-GFP and PompC(WT)-GFP. [http://2010.igem.org/Team:Tokyo_Tech/Project/wolf_coli/New_Series_of_PompC ...see more about PompC series]

;

UNIQ1bf67392673b4b26-partinfo-00000001-QINU

@@ Line 14: / Line 14: @@
 |width='60%' valign='top'|
 [[Image:Tokyotech_ompc_graph.jpg|thumb|center|400px|Figure 1. Induction of new'' OmpC'' series in high osmolarity medium ]]
-[https://parts.igem.org/wiki/index.php?title=Part:BBa_K395301 P''ompC(C)'']-GFP, [https://parts.igem.org/wiki/index.php?title=Part:BBa_K395302 P''ompC(CB)'']-GFP, [https://parts.igem.org/wiki/index.php?title=Part:BBa_K395303 P''ompC(CS1)'']-GFP and [https://parts.igem.org/wiki/index.php?title=Part:BBa_K395304 P''ompC(WT)'']-GFP 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) was used as a positive control and negative control respectively. <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 OD590 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>
+[https://parts.igem.org/wiki/index.php?title=Part:BBa_K395301 P''ompC(C)'']-GFP, [https://parts.igem.org/wiki/index.php?title=Part:BBa_K395302 P''ompC(CB)'']-GFP, [https://parts.igem.org/wiki/index.php?title=Part:BBa_K395303 P''ompC(CS1)'']-GFP and [https://parts.igem.org/wiki/index.php?title=Part:BBa_K395304 P''ompC(WT)'']-GFP 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) was 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 OD590 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 sucrose induction, 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]