Difference between revisions of "Part:BBa K395301:Experience"
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[[Image:Tokyotech_ompc_graph.jpg|thumb|center|400px|Figure 1. Induction of new'' OmpC'' series in high osmolarity medium ]] | [[Image:Tokyotech_ompc_graph.jpg|thumb|center|400px|Figure 1. Induction of new'' OmpC'' series in high osmolarity medium ]] | ||
Revision as of 11:42, 26 October 2010
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 placIq-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]
Applications of BBa_K395301
User Reviews
UNIQa1f906f9185d3369-partinfo-00000000-QINU UNIQa1f906f9185d3369-partinfo-00000001-QINU