Difference between revisions of "Part:BBa K5143027"
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<h1>Description</h1> | <h1>Description</h1> | ||
<p> | <p> | ||
| − | Our team has nicknamed this plasmid “<b>Venus/Ruby plasmid</b>”. Venus/Ruby plasmid is composed of the composite part Venus <a href="https://parts.igem.org/Part:BBa_K5143026" target="_blank">BBa_K5143026</a>, the | + | Our team has nicknamed this plasmid “<b>Venus/Ruby plasmid</b>”. Venus/Ruby plasmid is composed of the composite part Venus <a href="https://parts.igem.org/Part:BBa_K5143026" target="_blank">BBa_K5143026</a>, the composite part mruby2 <a href="https://parts.igem.org/Part:BBa_K5143021" target="_blank">BBa_K5143021</a> and the pUC57 backbone <a href="https://parts.igem.org/Part:BBa_K5143005" target="_blank">BBa_K5143005</a>. <br> |
| + | Venus/Ruby plasmid has been digest by XhoI restriction enzyme and the Venus/Ruby fragment has been transformed into the BY4741 <i> S. cerevisiae </i> strain. Then, this fragment has recombinated with BY4741 <i> S. cerevisiae </i> genome at the URA3 locus. This new strain has been tested for the expression of Venus and mRuby2 under the control of GAPpromoter and ADH1promoter respectively, by measuring fluorescence. It has also been tested for the secretion of the two different protein thanks to the alphafactor secretion signal peptide and the AGA2 pre signal peptide. <br> | ||
| + | These tests are essential to know in our project if the GAPpromoter works in our final construct <a href="https://parts.igem.org/Part:BBa_K5143025" target="_blank">BBa_K5143025</a> and if the fused peptides can be secreted thanks to the alphafactor. | ||
</p> | </p> | ||
Revision as of 08:03, 28 September 2024
pUC57-pGAP-AF_V1-Venus-ENO1_term-p_ADH1-AGA2-mRuby2-6xHis-TDH1_term
Description
Our team has nicknamed this plasmid “Venus/Ruby plasmid”. Venus/Ruby plasmid is composed of the composite part Venus BBa_K5143026, the composite part mruby2 BBa_K5143021 and the pUC57 backbone BBa_K5143005.
Venus/Ruby plasmid has been digest by XhoI restriction enzyme and the Venus/Ruby fragment has been transformed into the BY4741 S. cerevisiae strain. Then, this fragment has recombinated with BY4741 S. cerevisiae genome at the URA3 locus. This new strain has been tested for the expression of Venus and mRuby2 under the control of GAPpromoter and ADH1promoter respectively, by measuring fluorescence. It has also been tested for the secretion of the two different protein thanks to the alphafactor secretion signal peptide and the AGA2 pre signal peptide.
These tests are essential to know in our project if the GAPpromoter works in our final construct BBa_K5143025 and if the fused peptides can be secreted thanks to the alphafactor.
Construction
References