Difference between revisions of "Part:BBa K640001:Experience"
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This experience page is provided so that any user may enter their experience using this part.<BR>Please enter | This experience page is provided so that any user may enter their experience using this part.<BR>Please enter | ||
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===User Reviews=== | ===User Reviews=== | ||
| − | <!-- DON'T DELETE --><partinfo>BBa_K640001 StartReviews</partinfo> | + | 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<!-- DON'T DELETE --><partinfo>BBa_K640001 StartReviews</partinfo> |
<!-- Template for a user review | <!-- Template for a user review | ||
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Latest revision as of 20:03, 29 September 2011
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_K640001
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 partUNIQc1a3ebfe5e801445-partinfo-00000000-QINU UNIQc1a3ebfe5e801445-partinfo-00000001-QINU