Difference between revisions of "Part:BBa K2938015"
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[[File:Larva GFP.png|200px|thumb|left|Mosquitoes' larva that was fed with serratia marcescens 274 expressing plasmid with Cry11Aa and deGFP. confocal microscope (x10)]]
 
[[File:Larva GFP.png|200px|thumb|left|Mosquitoes' larva that was fed with serratia marcescens 274 expressing plasmid with Cry11Aa and deGFP. confocal microscope (x10)]]
 
[[File:Cry11Aa_Western.png|200px|thumb|right|Western Blot of Bacteria expressing Cry11Aa and deGFP – proof of Cry11Aa expression]]
 
[[File:Cry11Aa_Western.png|200px|thumb|right|Western Blot of Bacteria expressing Cry11Aa and deGFP – proof of Cry11Aa expression]]
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<partinfo>BBa_K2938016 SequenceAndFeatures</partinfo>
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===Functional Parameters===
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Revision as of 10:16, 12 October 2019


Cry11Aa (Strep tag) + deGFP Device

PR1 - RBS - Cry11Aa(Strep tag) - RBS - deGFP - T500 Terminator

This plasmid was planned on a pBEST plasmid backbone. It contains larvicidal activity, as it containing toxic subunit of the bacillus thuringiensis israelensis (BTI) pBtoxis plasmid which is toxic to mosquitoes larva.

The plasmid was constructed using gibson assembly, then was incorporated into E.coli BL21 and DH5a using Heat shock, and into serratia marcescens 274 using electroporation.


Characterization

Western of Cry11Aa subunit with anti-Strep antibody. proving the translation of the toxin subunit on the plasmid.

and confocal microscope photo showing our transgenic bacteria presence in the mosquitoes larva

Mosquitoes' larva that was fed with serratia marcescens 274 expressing plasmid with Cry11Aa and deGFP. confocal microscope (x10)
Western Blot of Bacteria expressing Cry11Aa and deGFP – proof of Cry11Aa expression



Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1870
    Illegal EcoRI site found at 2426
    Illegal XbaI site found at 2116
    Illegal XbaI site found at 2761
    Illegal SpeI site found at 982
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1870
    Illegal EcoRI site found at 2426
    Illegal NheI site found at 56
    Illegal NheI site found at 105
    Illegal SpeI site found at 982
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1870
    Illegal EcoRI site found at 2426
    Illegal XhoI site found at 3494
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1870
    Illegal EcoRI site found at 2426
    Illegal XbaI site found at 2116
    Illegal XbaI site found at 2761
    Illegal SpeI site found at 982
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1870
    Illegal EcoRI site found at 2426
    Illegal XbaI site found at 2116
    Illegal XbaI site found at 2761
    Illegal SpeI site found at 982
  • 1000
    COMPATIBLE WITH RFC[1000]