Difference between revisions of "Part:BBa K640000:Experience"

Latest revision as of 03:50, 29 September 2011

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Applications of BBa_K640000

User Reviews

Preliminary characterization data was collected for a microalgae construct containing both the Hsp70A/RbcS2 promoter attached to the algal luciferase. Chlamydomonas reibhrdtii cells were transformed with a plasmid DNA of a construct containing the Hsp70A/RbcS2 promoter upstream of luciferase. This was done using our optimized glass bead method. Following transformation, cells were incubated at various temperatures (4°C, 21°C, 30°C, 37°C and 50°C) as well as in the dark for 1 hour. After this, luminescence and OD was measured (at 595 nm) using a Victor. When CPS was corrected for OD, luciferase activity was seen for cells incubated at 37°C and 50°C. Although this was higher than baseline levels, is was not very significant. This is likely due to the very small number of algal cells. This indicates that our luciferase seems to be working, and our promoter is also working, however we cannot make any statements about how inducible it may be. Further characterization will be needed for this part UNIQ730a4cf971c18956-partinfo-00000000-QINU UNIQ730a4cf971c18956-partinfo-00000001-QINU

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 ===User Reviews===
+Preliminary characterization data was collected for a microalgae construct containing both the Hsp70A/RbcS2 promoter attached to the algal luciferase. Chlamydomonas reibhrdtii cells were transformed with a plasmid DNA of a construct containing the Hsp70A/RbcS2 promoter upstream of luciferase. This was done using our optimized glass bead method. Following transformation, cells were incubated at various temperatures (4°C, 21°C, 30°C, 37°C and 50°C) as well as in the dark for 1 hour. After this, luminescence and OD was measured (at 595 nm) using a Victor. When CPS was corrected for OD, luciferase activity was seen for cells incubated at 37°C and 50°C. Although this was higher than baseline levels, is was not very significant. This is likely due to the very small number of algal cells. This indicates that our luciferase seems to be working, and our promoter is also working, however we cannot make any statements about how inducible it may be. Further characterization will be needed for this part
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