Difference between revisions of "Part:BBa K4800013"
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<p style="font-size: 180%; font-weight: bold;">Build: | <p style="font-size: 180%; font-weight: bold;">Build: | ||
− | <p>< img src="https://static.igem.wiki/teams/4800/wiki/parts/ycjq-sgrna.png" height="80%"></p > | + | <p><img src="https://static.igem.wiki/teams/4800/wiki/parts/ycjq-sgrna.png" width="80%" height="80%"></p> |
<div align="center"> | <div align="center"> | ||
<strong>Fig.1 YcjQ sgRNA plasmid</strong> | <strong>Fig.1 YcjQ sgRNA plasmid</strong> | ||
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<p style="font-size: 180%; font-weight: bold;">Test: | <p style="font-size: 180%; font-weight: bold;">Test: | ||
− | <p>< img src="https://static.igem.wiki/teams/4800/wiki/parts/yciq-sgrna-result.png"></p > | + | <p><img src="https://static.igem.wiki/teams/4800/wiki/parts/yciq-sgrna-result.png" width="80%" height="80%"></p> |
<div align="center"> | <div align="center"> | ||
− | <strong>Fig.2 YcjQ sgRNA fermentation result</strong> | + | <strong>Fig.2 YcjQ sgRNA fermentation result</strong> |
</div> | </div> | ||
<p>We made the knockout strain into competent cells, and then PRSFDuet-YciA-sfp-MmCAR-YahK and PTrc99a-davB-davA-GabT were transferred into KA30 competent cells knocked out of the ycjQ gene by electrotransfer. After verifying the success of plasmid transfer, they were plated into 5 mL of KA30 seed medium at 37°C and transferred into shake flasks after 20 h. Fermentation was carried out at 37°C and 200 rpm.</p > | <p>We made the knockout strain into competent cells, and then PRSFDuet-YciA-sfp-MmCAR-YahK and PTrc99a-davB-davA-GabT were transferred into KA30 competent cells knocked out of the ycjQ gene by electrotransfer. After verifying the success of plasmid transfer, they were plated into 5 mL of KA30 seed medium at 37°C and transferred into shake flasks after 20 h. Fermentation was carried out at 37°C and 200 rpm.</p > |
Revision as of 07:45, 6 October 2023
YcjQ sgRNA
SgRNA, or small guide RNA, is an important component of the CRISPR knockdown system. The N20 sequence of the ycjQ SgRNA is derived from the ycjQ gene and can be localised to the ycjQ gene in the KA30 genome to assist the Cas9 protein in silencing the gene.
Build:
The N20 sequence of the YcjQ gene was found on the CHOPCHOP website, designed on primers and the corresponding SgRNA was constructed by IPCR from the original SgRNA plasmid.
Test:
We made the knockout strain into competent cells, and then PRSFDuet-YciA-sfp-MmCAR-YahK and PTrc99a-davB-davA-GabT were transferred into KA30 competent cells knocked out of the ycjQ gene by electrotransfer. After verifying the success of plasmid transfer, they were plated into 5 mL of KA30 seed medium at 37°C and transferred into shake flasks after 20 h. Fermentation was carried out at 37°C and 200 rpm.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]