Difference between revisions of "Part:BBa K2938014"
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===Usage and Biology===
 
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Revision as of 10:16, 12 October 2019


Cry4Ba (HA tag) + deGFP Device

PR1 - RBS - Cry4Ba (HA 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 Cry4Ba subunit with anti-HA antibody. proving the translation of the toxin subunit on the plasmid.

Western Blot of Bacteria expressing Cry4Ba and deGFP – proof of Cry4Ba 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]