Difference between revisions of "Part:BBa K3930022"

 
 
(12 intermediate revisions by 3 users not shown)
Line 2: Line 2:
 
__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K3930022 short</partinfo>
 
<partinfo>BBa_K3930022 short</partinfo>
 +
<span class='h3bb'>Sequence and Features</span>
 +
<partinfo>BBa_K3930022 SequenceAndFeatures</partinfo>
  
−
Integrative locus XII-4 DOWN of S. cerevisiae genome comes from the EasyClone-MarkerFree kit (Jessop-Fabre et al.,2016). This part needs to flank the insertion in 3', and must be used with the integrative locus (BBa_K39300021) XII-4 up part in 5'.
+
<html>
 +
 
 +
<h2>Introduction</h2>
 +
<p>The sequences to target the locus XII-4 (Chr XII:830227..831248) of <i>S. cerevisiae</i> genome come from the EasyClone-MarkerFree kit (Jessop-Fabre et al., 2016).
 +
This part is flanking the insert in 3' and must be used with the <a href="https://parts.igem.org/Part:BBa_K3930021" class="pr-0" target="_blank">(BBa_K3930021)</a> XII-4 up part in 5'.</p>
 +
<h2>Results</h2>
 +
 
 +
<h3>Integration of part <a href="https://parts.igem.org/Part:BBa_K3930003" class="pr-0" target="_blank">(BBa_K3930003)</a> into the yeast genome at locus XII-4</h3>
 +
<p> The part <a href="https://parts.igem.org/Part:BBa_K3930003" class="pr-0" target="_blank">(BBa_K3930003)</a> was linearized and transformed into the <i>S. cerevisiae</i> LycoYeast strain according to the Takara yeast transformation protocol, with 5 µg of DNA. The insert is flanked by parts <a href="https://parts.igem.org/Part:BBa_K3930021" class="pr-0" target="_blank">(BBa_K3930021)</a> and (BBa_K3930022). Figure 1 shows the electrophoresis gel of colony PCR to verify integrants genotype (clone 2 is OK).</p>
 +
<br>
 +
<br>
 +
<div class="center">
 +
    <div class="thumb tnone">
 +
        <div class="thumbinner" style="width:50%;">
 +
            <a href="https://2021.igem.org/wiki/images/a/a9/T--Toulouse_INSA-UPS--2021_fig23.violette.png" class="image">
 +
                <img alt="" src="https://2021.igem.org/wiki/images/a/a9/T--Toulouse_INSA-UPS--2021_fig23.violette.png" width="100%" height=auto class="thumbimage" /></a>                  <div class="thumbcaption">
 +
                <div class="magnify">
 +
                    <a href="https://2021.igem.org/wiki/images/a/a9/T--Toulouse_INSA-UPS--2021_fig23.violette.png" class="internal" title="Enlarge"></a>
 +
                </div>
 +
                <b>Figure 1: </b> <b> Integration of pVIOLETTE insert in the LycoYeast genome at locus XII-4</b>
 +
                <p>pVIOLETTE insert integration at locus XII-4 was checked by PCR and the result was visualized on EtBr stained agarose electrophoresis gel. A theoretical gel is presented on the right (note that a different ladder is presented on the theoretical gel).</p>
 +
            </div>
 +
        </div>
 +
    </div>
 +
</div>
 +
<br>
 +
<p><b>The sequence XII-4 down (BBa_K3930022) coupled with the XII-4 up part <a href="https://parts.igem.org/Part:BBa_K3930021" class="pr-0" target="_blank">(BBa_K3930021)</a> is functional to target integration at the XII-4 locus under these experimental conditions.</b><p>
 +
<br>
 +
<h2>References</h2>
 +
<ol>
 +
    <i>
 +
    <li>Chen X, Shukal S, Zhang C. 2019. Integrating Enzyme and Metabolic Engineering Tools for Enhanced α-Ionone Production. J Agric Food Chem. 67(49):13451–13459. doi:10.1021/acs.jafc.9b00860.</li>
 +
</i>
 +
</ol>
 +
</html>
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here
Line 9: Line 45:
  
 
<!-- -->
 
<!-- -->
−
<span class='h3bb'>Sequence and Features</span>
 
−
<partinfo>BBa_K3930022 SequenceAndFeatures</partinfo>
 
  
  

Latest revision as of 21:19, 16 October 2021


Down Integrative sequence to target locus XII-4 of Saccharomyces cerevisiae genome 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]

Introduction

The sequences to target the locus XII-4 (Chr XII:830227..831248) of S. cerevisiae genome come from the EasyClone-MarkerFree kit (Jessop-Fabre et al., 2016). This part is flanking the insert in 3' and must be used with the (BBa_K3930021) XII-4 up part in 5'.

Results

Integration of part (BBa_K3930003) into the yeast genome at locus XII-4

The part (BBa_K3930003) was linearized and transformed into the S. cerevisiae LycoYeast strain according to the Takara yeast transformation protocol, with 5 µg of DNA. The insert is flanked by parts (BBa_K3930021) and (BBa_K3930022). Figure 1 shows the electrophoresis gel of colony PCR to verify integrants genotype (clone 2 is OK).



Figure 1: Integration of pVIOLETTE insert in the LycoYeast genome at locus XII-4

pVIOLETTE insert integration at locus XII-4 was checked by PCR and the result was visualized on EtBr stained agarose electrophoresis gel. A theoretical gel is presented on the right (note that a different ladder is presented on the theoretical gel).


The sequence XII-4 down (BBa_K3930022) coupled with the XII-4 up part (BBa_K3930021) is functional to target integration at the XII-4 locus under these experimental conditions.


References

  1. Chen X, Shukal S, Zhang C. 2019. Integrating Enzyme and Metabolic Engineering Tools for Enhanced α-Ionone Production. J Agric Food Chem. 67(49):13451–13459. doi:10.1021/acs.jafc.9b00860.
    2.