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| | The g-ilvB is a guide RNA that can form a complex with Cas 9 in E.coli cicc 20905. It is a specific RNA sequence (around 20 bp) that recognizes the ilvB gene and directs the Cas 9 protein there for gene knocking out. | | The g-ilvB is a guide RNA that can form a complex with Cas 9 in E.coli cicc 20905. It is a specific RNA sequence (around 20 bp) that recognizes the ilvB gene and directs the Cas 9 protein there for gene knocking out. |
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| − | <!-- Add more about the biology of this part here
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| − | <h2><b>Nature function description【Usage and Biology】</b>></h2>
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| − | <p>The g-rhtA is a guide RNA that can form a complex with Cas 9 in E.coli cicc 20905. It is a specific RNA sequence
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| − | (around 20 bp) that recognizes the rhtA gene and directs the Cas 9 protein there for gene knocking out.</p>
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| − | <h2><b>long description【Characterization】</b></h2>
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| − | <p>rhtA gene encodes a strong threonine exporter and transfers threonine out of the cell. The gene rhtA knockout
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| − | resulted in the reduction of extracellular secretion of threonine so there are more amount of threonine can be
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| − | kept inside the cell for the further production of a-KB. We design the pTarget plasmid that carrying specific
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| − | gRNA sequence which can identity the rhtA gene, then we obtained donorDNA through two rounds of PCR. The
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| − | donorDNA was used for homologous recombination with genomic DNA. We then put this pTarget plasmid and donor DNA
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| − | into AIS-0 strains that has already carried pEcCas plasmid for the CRISPR-CAS 9 knockout experiment. (We
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| − | referred to the experimental procedures published by Qi Li, Bingbing Sun, et al. in 2020) Through the results of
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| − | colony PCR and gene sequencing, we confirmed the successful knockout of rhtA. </p>
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| − | <img src=https://static.igem.wiki/teams/4808/wiki/parts1.png alt="" class="w-100">
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| − | <!-- -->
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| − | <span class='h3bb'>Sequence and Features</span>
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| − | <partinfo>BBa_K4808005 SequenceAndFeatures</partinfo>
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Revision as of 10:15, 10 October 2023
g-ilvB
The g-ilvB is a guide RNA that can form a complex with Cas 9 in E.coli cicc 20905. It is a specific RNA sequence (around 20 bp) that recognizes the ilvB gene and directs the Cas 9 protein there for gene knocking out.
