Difference between revisions of "Part:BBa K3365059:Design"

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===Design Notes===
 
===Design Notes===
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We choose additional antibiotic resistance genes that was not used before, kanamycin resistant gene as the alternatives to <i>rpoZ</i>, so that we can make a preliminary judgment with the medium containing kanamycin. Only the baterial which was knocked out <i>rpoZ</i> can survive in this medium while others cannot survive.
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We choose additional antibiotic resistance genes that was not used before, kanamycin resistant gene as the alternatives to <i>rpoZ</i>, so that we can make a preliminary judgment with the medium containing kanamycin. Only the bacteria whose genome was knocked out <i>rpoZ</i> can survive in kanamycin resistant medium while others not.
  
  

Latest revision as of 12:47, 27 October 2020


kanamycin resistant gene with homologous arms


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]


Design Notes

We choose additional antibiotic resistance genes that was not used before, kanamycin resistant gene as the alternatives to rpoZ, so that we can make a preliminary judgment with the medium containing kanamycin. Only the bacteria whose genome was knocked out rpoZ can survive in kanamycin resistant medium while others not.


Source

We get the sequence of homologous arm at both ends of rpoZ from reference and get the part of kanamycin resistant gene from pET28a contained kanamycin resistance through PCR.

References

[1]David Bikard,Wenyan Jiang,Poulami Samai,Ann Hochschild,Feng Zhang,Luciano A. Marraffini1.Programmable repression and activation of bacterial gene expression using an engineered CRISPR-Cas system[J]. Nucleic Acids Research,2013,41(15):7429-7437.