Difference between revisions of "Part:BBa J63006:Experience"
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==Characterization by British Columbia iGEM 2011== | ==Characterization by British Columbia iGEM 2011== | ||
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+ | ===Fluorescence Analysis of GFP expression as regulated by GPD and GAL Promoters=== | ||
The GPD promoter (<partinfo>BBa_K517000</partinfo>), GAL promoter (<partinfo>BBa_K517001</partinfo>) and GAL1 promoter with Kozak sequence (<partinfo>BBa_J63006</partinfo>) as characterized by their regulation of the expression of a GFP reporter. | The GPD promoter (<partinfo>BBa_K517000</partinfo>), GAL promoter (<partinfo>BBa_K517001</partinfo>) and GAL1 promoter with Kozak sequence (<partinfo>BBa_J63006</partinfo>) as characterized by their regulation of the expression of a GFP reporter. | ||
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[[Image:Ubcigem2011Promoterimages.jpg | frame | center | '''Fluorescence Analysis of GFP expression as regulated by GPD and GAL Promoters:''' ''S. cerevisiae'' yeast strains containing either the GPD-GFP, GAL-GFP or GAL+Kozak-GFP construct were cultured overnight at 30 degrees Celsius in either YPD (dextrose), SC-raffinose or SC-galactose media. These were diluted 1 in 10 in their respective media and grown for 3 hours at 30 degrees Celsius into log phase. The cells were then spun down and samples were collected at 0 hours. The remaining cells were resuspended in SC-galactose media and left to grow at 30 degrees Celsius for 3 hours. The cells were spun down again and samples were collected at this 3 hour time point. The samples were fixed in paraformaldehyde and visualized under a fluorescence microscope under the GFP and DIC settings. Acquired images were then color-combined with red representing DIC and green representing GFP.]] | [[Image:Ubcigem2011Promoterimages.jpg | frame | center | '''Fluorescence Analysis of GFP expression as regulated by GPD and GAL Promoters:''' ''S. cerevisiae'' yeast strains containing either the GPD-GFP, GAL-GFP or GAL+Kozak-GFP construct were cultured overnight at 30 degrees Celsius in either YPD (dextrose), SC-raffinose or SC-galactose media. These were diluted 1 in 10 in their respective media and grown for 3 hours at 30 degrees Celsius into log phase. The cells were then spun down and samples were collected at 0 hours. The remaining cells were resuspended in SC-galactose media and left to grow at 30 degrees Celsius for 3 hours. The cells were spun down again and samples were collected at this 3 hour time point. The samples were fixed in paraformaldehyde and visualized under a fluorescence microscope under the GFP and DIC settings. Acquired images were then color-combined with red representing DIC and green representing GFP.]] | ||
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Characterization by British Columbia iGEM 2011
Fluorescence Analysis of GFP expression as regulated by GPD and GAL Promoters
The GPD promoter (BBa_K517000), GAL promoter (BBa_K517001) and GAL1 promoter with Kozak sequence (BBa_J63006) as characterized by their regulation of the expression of a GFP reporter.