A SAI AIA , , TH THE PIN INK FORC RCE A GA INST M AL ARIA GAIN ALARIA
Watch our animated movie which explains our project : http://2010.igem.org/Team:EPF_Lausanne/Humanpractices_Movie
p2 p25 5 / p / p28
p2 p25 5 / p / p28
p2 p25 5 / p / p28
p25 / p p2 / p28 Im Immun unotox otoxin in
Yoshida et al, Molecular and Biochemical Prasitology, 2000
pelB FLAG Tag Heavy chain linker Light chain Shiva HIS Tag Pore-forming Antibody Toxin fragment
1 st goal Find a way to deliver these proteins to the mosquito’s gut!
ASAIA ADDRESS : mos osquito's ito's intestin estinal al tra ract SPECIAL CHARACTERISTIC : pink! k! ADDITIONAL INFO : speci ecific fic or origi gin RECENTLY FRIEND WITH : E. E.col oli
Asaia is a commensal bacterium of mosquitoes Male terminal portion of mitgut Female terminal portion of mitgut Favia et al., PNAS 2007
Horizontally Vertically
Our project
Find a way to deliver these proteins to the mosquito’s gut!
2 nd goal Establish Asaia as a new chassis for iGEM
Asaia growth characterization pH5 Best conditions : 30°C, pH5
Asaia GFP in Drosophila 3 hours after infection Observation of fluorescent Asaia in the Drosophila's gut
Crotti et al., AEM Acetic acid bacteria, new emerging symbionts of insects, 2010
Asaia origin Extracted from pHM2-GFP Plasmid Compatible with E. coli
Asaia Asaia E. coli origin origin origin
Characterized Asaia growth Characterized Asaia’s antibiotic resistance spectrum Tested Asaia infection in Drosophila Generated tech sheets for working with Asaia: growth, competent cells, transformation, and cloning
Establish Asaia as a new chassis
3 rd goal Construct and characterize BioBricks
Lysate Supernatant
Construct and characterize BioBricks
Human practices
Express p25, p28, and the Immunotoxin in Asaia Test in vivo (Institute Pasteur) Introduce the engineered Asaia strain in the wild
Acknowledgements
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