Difference between revisions of "Part:BBa K3930021"
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<h2>Introduction</h2> | <h2>Introduction</h2> | ||
− | <p> | + | <p>The sequence 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 | + | This part is flanking the insert in 5' and must be used with the <a href="https://parts.igem.org/Part:BBa_K3930022" class="pr-0" target="_blank">(BBa_K3930022)</a> XII-4 up part in 3'.</p> |
<h2>Results</h2> | <h2>Results</h2> | ||
− | <h3>Integration of | + | <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 (BBa_K3930003) 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 construction is flanked by | + | <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 construction is flanked by parts (BBa_K39300021) and <a href="https://parts.igem.org/Part:BBa_K3930022" class="pr-0" target="_blank">(BBa_K3930022)</a>. Figure 1 shows the electrophoresis gel of colony PCR to verify integrants genotype (clone 2 is OK).</p> |
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− | <a href=" | + | <a href="https://2021.igem.org/wiki/images/a/a9/T--Toulouse_INSA-UPS--2021_fig23.violette.png" class="image"> |
− | <img alt="" src="/wiki/images/ | + | <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"> |
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− | <a href=" | + | <a href="https://2021.igem.org/wiki/images/a/a9/T--Toulouse_INSA-UPS--2021_fig23.violette.png" class="internal" title="Enlarge"></a> |
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− | <b>Figure 1: </b> <b> Integration of pVIOLETTE in the LycoYeast genome</b> | + | <b>Figure 1: </b> <b> Integration of pVIOLETTE insert in the LycoYeast genome at locus XII-4</b> |
− | <p>pVIOLETTE was checked | + | <p>pVIOLETTE insert integration was checked by PCR visualised 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> |
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− | <p><b>The | + | <p><b>The integrative locus XII-4 up (BBa_K3930021) coupled with the integrative locus <a href="https://parts.igem.org/Part:BBa_K3930022" class="pr-0" target="_blank">(BBa_K3930022)</a> XII-4 down part is functional to target integration at the XII-4 locus under those experimental conditions.</b><p> |
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<h2>References</h2> | <h2>References</h2> |
Latest revision as of 21:22, 16 October 2021
Up integrative sequence to target locus XII-4 of Saccharomyces cerevisiae genome
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 73
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Introduction
The sequence 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 5' and must be used with the (BBa_K3930022) XII-4 up part in 3'.
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 construction is flanked by parts (BBa_K39300021) and (BBa_K3930022). Figure 1 shows the electrophoresis gel of colony PCR to verify integrants genotype (clone 2 is OK).
The integrative locus XII-4 up (BBa_K3930021) coupled with the integrative locus (BBa_K3930022) XII-4 down part is functional to target integration at the XII-4 locus under those experimental conditions.
References
- 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.