Difference between revisions of "Part:BBa K4808005"

 
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===Usage and Biology===
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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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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>

Revision as of 10:02, 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.

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]