Difference between revisions of "Team:Uppsala/Improve"

Revision as of 18:13, 31 October 2017

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Our project was in a sense, a continuation of Uppsala 2013 and Slovenia 2010 team's legacy. We have worked with the Slovenia 2010 strain, and integrated the genes into the chromosome to stabilize production. We improved the crocin biosynthetic pathway that Uppsala 2013 had problems with by identifying and working with the correct genes in the pathway i.e CaCCD2 and CsADH2946. We have done codon optimization of all the enzyme sequences and created new biobricks with them. Another improvement we made was to add different promoters to the biobrick. In all of our enzymes, we have worked with both constitutive and inducible promoters. Since our enzymes were poorly characterized , we added a his-tag to them to make the purification more achievable. The addition of his-tag was done after consulting the structural models of the enzymes. For example, BBa_K2423008 is an improved version of BBa_K1033112 and a slightly modified version of BBa_K2423002 (different promoter and RBS).

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        <div class="header" style="margin-top:100px;">Improvement</div>
        <img src="https://static.igem.org/mediawiki/2017/3/30/Results_line_under_title.svg" style="margin: auto;">
-       <div class="textbox">Our project was in a sense,a continuation of Uppsala 2013 and Slovenia 2010 team's legacy. We have worked with the Slovenia 2010 strain and transported it to the chromosome to stabilize production. We improved the crocin  biosynthethetic pathway that Uppsala 2013 had problems with by identifying and working with the correct genes in the pathway i.e CaCCD2 and CsADH2946. We have done codon optimization of all the enzyme sequences and created new biobricks with them. Another improvement we made was to add different promoters to the biobrick. In all of our enzymes, we have worked with both constitutive and inducible promoters. Since our enzymes were poorly characterzed , we added a his-tag to them to make the purification more achievable. The addition of his-tag was done after consulting the structural models of the enzymes.  For example, <a href="http://parts.igem.org/Part:BBa_K2423008">BBa_K2423008</a> is an improved version of <a href="http://parts.igem.org/Part:BBa_K1033112">BBa_K1033112</a> and a slightly modified version of <a href="http://parts.igem.org/Part:BBa_K2423002">BBa_K2423002</a> (different promoter and RBS).
+       <div class="textbox">Our project was in a sense, a continuation of Uppsala 2013 and Slovenia 2010 team's legacy. We have worked with the Slovenia 2010 strain, and integrated the genes into the chromosome to stabilize production. We improved the crocin  biosynthetic pathway that Uppsala 2013 had problems with by identifying and working with the correct genes in the pathway i.e CaCCD2 and CsADH2946. We have done codon optimization of all the enzyme sequences and created new biobricks with them. Another improvement we made was to add different promoters to the biobrick. In all of our enzymes, we have worked with both constitutive and inducible promoters. Since our enzymes were poorly characterized , we added a his-tag to them to make the purification more achievable. The addition of his-tag was done after consulting the structural models of the enzymes. For example, <a href="http://parts.igem.org/Part:BBa_K2423008">BBa_K2423008</a> is an improved version of <a href="http://parts.igem.org/Part:BBa_K1033112">BBa_K1033112</a> and a slightly modified version of <a href="http://parts.igem.org/Part:BBa_K2423002">BBa_K2423002</a> (different promoter and RBS).
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