Difference between revisions of "Part:BBa K2294007:Experience"
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Results: <br> | Results: <br> | ||
<img src="https://static.igem.org/mediawiki/2017/c/c9/T--BOKU-Vienna--Promoterstrenght_comparisson.png" alt="promoterstrength" /> | <img src="https://static.igem.org/mediawiki/2017/c/c9/T--BOKU-Vienna--Promoterstrenght_comparisson.png" alt="promoterstrength" /> | ||
| − | <br> | + | <br><br> |
| − | There is no significant difference between the different galactose concentrations which were investigated. The galpromoter combined with a Kozak sequnce showed the highes expression levels of GFP. It is about twice compared to the gal promoter without a Kozak sequence and three times higher compared to the pTDH3 promoter of <i>Saccharomyces cerevisiae </i>. | + | There is no significant difference between the different galactose concentrations which were investigated. The galpromoter combined with a Kozak sequnce showed the highes expression levels of GFP. It is about twice compared to the gal promoter without a Kozak sequence and three times higher compared to the pTDH3 promoter of <i>Saccharomyces cerevisiae </i>. The clones expressing GFP under the control of the consitutive TDH3 promoter were cultivated in YP + 2% glucose. |
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Characterization by BOKU-Vienna 2017
For characterization the synGAL promoter was combined with GFP and the CYC1 terminator and afterwards integrated into the ura3 gene of Saccharomyces cerevisiae . The GFP fluorescence was measured with a flow cytometer after inducing with 3 different galactose concentrations (0,5%, 1% and 2%).
The cultivation was performded in 24 deepwell plates using 2 mL YP media. As controls a wildtype and a clone expressing GFP under the constituve pTDH3 promoter were cultivated. In addition, the synGAL promoter without a Kozak sequence was investigated.
Results:
<img src="
" alt="promoterstrength" />
There is no significant difference between the different galactose concentrations which were investigated. The galpromoter combined with a Kozak sequnce showed the highes expression levels of GFP. It is about twice compared to the gal promoter without a Kozak sequence and three times higher compared to the pTDH3 promoter of Saccharomyces cerevisiae . The clones expressing GFP under the control of the consitutive TDH3 promoter were cultivated in YP + 2% glucose.