Part:BBa_K509002:Experience
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Applications of BBa_K509002
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UNIQa0a2c08fc34d6ed8-partinfo-00000000-QINU UNIQa0a2c08fc34d6ed8-partinfo-00000001-QINU
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iGEM Concordia 2014 |
Group name: iGEM Concordia 2014 Author: Victor Yuan Summary: We used this promoter by constructing it into a GFP-NLS cassette. It precedes a GFP-NLS coding sequence, which ends in the PSAD terminator too (BBa_K509003, BBa_K1547010), and we transformed into chlamydomonas reinhardtii. Additionally, this entire cassette was submitted as a composite part (BBa_K1547001) on behalf of the iGEM Concordia 2014 team. We confirmed that the cassette was integrated by performing a TAQ PCR on genomic DNA extract of transformants, using specific cassette-specific primers. We were able to visualize expression of GFP under fluorescent microscopy and very intense bands relative to the other promoter we tested, HSP70/RBCS2m, were seen. We conclude that the PSAD promoter will express a sufficient amount of GFP to be observed under fluorescent microscopy, and further characterization could include qualitative analysis such as using FLOW cytometry (although adjustment for chloroplast interference should be considered).
GFP-NLS expression using PSAD promoter visualized under fluorescent microscopy
Wildtype c. reinhardtii cells showing a lack of bright regions
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