Difference between revisions of "Part:BBa K2365508"
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<partinfo>BBa_K2365508 parameters</partinfo> | <partinfo>BBa_K2365508 parameters</partinfo> | ||
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| + | [[File:Change.png|300px|center]] | ||
| + | The 579bp fragment was cloned under the GAL1 promoter, followed by CYC1 terminator. The pYES2 vector is designed for native expression of protein of interest in S. cerevisiae. It contains the URA3 gene for selection in yeast and 2µ origin for high-copy maintenance. After verifying the right constructs by restriction enzyme digestions and sequencing, the plasmids were transformed into S. cerevisiae SEY6210. | ||
| + | [[File:BAX CCK.jpeg|400px|center]] | ||
| + | CCK-8 testing | ||
| + | Method: | ||
| + | Cells were added into 96-well microplates. Then CCK8 solution were added to the wells and the plates were incubated for additional 4 h at 30°C. Optical density was measured using a microplate reader at a wavelength of 450 nm. | ||
Latest revision as of 00:14, 2 November 2017
Bax induced part
This part is used to as the key part of our biosafety device. Coding sequence of Bax protein is linked to the Gal1 promotor. It could be inserted to your device as kill switch when your chassis fungal is yeast(in the most broadly yeast).
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 900
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 70
- 1000COMPATIBLE WITH RFC[1000]
The 579bp fragment was cloned under the GAL1 promoter, followed by CYC1 terminator. The pYES2 vector is designed for native expression of protein of interest in S. cerevisiae. It contains the URA3 gene for selection in yeast and 2µ origin for high-copy maintenance. After verifying the right constructs by restriction enzyme digestions and sequencing, the plasmids were transformed into S. cerevisiae SEY6210.
CCK-8 testing Method: Cells were added into 96-well microplates. Then CCK8 solution were added to the wells and the plates were incubated for additional 4 h at 30°C. Optical density was measured using a microplate reader at a wavelength of 450 nm.