Difference between revisions of "Part:BBa K3221205"
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===Usage and Biology===
 
===Usage and Biology===
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The yamilCP was improved from amilCP(Part: BBa_K592009) , which is a blue chromoprotein gene from <i>Acropora millepora</i> submitted by Team Uppsala-Sweden 2011. <br>
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<p>The yamilCP was improved from amilCP(Part: BBa_K592009) [https://parts.igem.org/Part:BBa_K592009 BBa_K592009], which is a blue chromoprotein gene from <i>Acropora millepora</i> submitted by Team Uppsala-Sweden 2011.</p>
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We expected amilCP to be expressed efficiently in <i>S. cerevisiae</i>, so we chose amilCP to be improved. Consequently, we processed codon optimization on amilCP and added a consensus sequence for its usage in <i>S. cerevisiae</i>.  
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We processed codon optimization and added a consensus sequence(AAAAAA) on the original amilCP sequence and obtained the sequence of yamilCP.  
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<p>We expected amilCP to be expressed efficiently in <i>S. cerevisiae</i>, so we chose amilCP to be improved. We processed codon optimization on original amilCP sequence and added a consensus sequence (AAAAAA) for its usage in <i>S. cerevisiae</i>. And the new sequence was named yamilCP.</p>
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<p>We respectively cloned the amilCP and yamilCP on the vector of pYES2-NTA.There are the maps of pYES2-amilCP and pYES2-yamilCP (Figure1 and Figure 2). And we verified the construction of them by double restriction enzymes digestion and sequencing (Figure 3).</p>
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<p>Then we transformed pYES2-amilCP and pYES2-yamilCP into <i>S. cerevisiae</i> respectively and successfully observed the expression of blue chromoprotein (Figure 4).</p>
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<p>To test the expression of pYES2-amilCP and pYES2-yamilCP in <i>S. cerevisiae</i>, we transformed them into <i>S. cerevisiae</i> BY4741 and used galactose to induce blue chromoprotein expression. After 10 days, we observed blue colony on the pYES2-yamilCP plate at first and there was no blue colony on the pYES2-amilCP plate, which confirm that yamilCP express more quickly than amilCP (Figure 5). It shows that our improvement is successful.</p>
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<p>As the result, the improved yamilCP was successfully expressed, and the expression of yamilCP is more quickly than that of amilCP in <i>S. cerevisiae</i>. </p>
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Revision as of 10:31, 19 October 2019


YamilCP is a CDS of blue chromoprotein in S. cerevisiae.

YamilCP is a CDS of chromoprotein which can exhibit strong blue color in S. cerevisiae. It was optimized for S. cerevisiae on the basis of amilCP(BBa_K592009) which is encouraged by Team Uppsala-Sweden 2011. It acts as a reporter in our project to determine the function of Gal1 promoter (pGAL1) and Met3 promoter (pMet3).


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]