Difference between revisions of "Part:BBa K2688027:Design"

 
 
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===Design Notes===
 
===Design Notes===
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In pSB1C3-LEE5*Δtir-gfp, the tir fragment was deleted so that the gfp start codon is at the position of the former tir start codon. Our results indicate that gfp expression is improved when this fragment is removed compared to the parental pSB1C3- LEE5*-gfp plasmid, and that H-NS protein is still able to control the remaining LEE5 region. Of note, Ler proteins are able to activate LEE5 promoter.
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In pSB1C3-LEE5*Δtir-gfp, the tir fragment was deleted so that the gfp start codon is at the position of the former tir start codon, which yielded BBa_K2688017. The introduced point mutation was then PCR corrected, yielding this biobrick.
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Our results indicate that gfp expression is improved when this fragment is removed compared to the parental pSB1C3- LEE5*-gfp plasmid, and that H-NS protein is still able to control the remaining LEE5 region. Of note, Ler proteins are able to activate LEE5 promoter.
  
  

Latest revision as of 16:16, 28 September 2018


LEE5_GFP_native_ΔTir


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
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 50
    Illegal BsaI.rc site found at 1003


Design Notes

In pSB1C3-LEE5*Δtir-gfp, the tir fragment was deleted so that the gfp start codon is at the position of the former tir start codon, which yielded BBa_K2688017. The introduced point mutation was then PCR corrected, yielding this biobrick.

Our results indicate that gfp expression is improved when this fragment is removed compared to the parental pSB1C3- LEE5*-gfp plasmid, and that H-NS protein is still able to control the remaining LEE5 region. Of note, Ler proteins are able to activate LEE5 promoter.



Source

LEE5 is a topic of research at the host lab, which provided us with the source plasmid pKK-LEE5-gfp.

References