Team:Trieste/notebook3

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Week 3

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Suicide System

We inoculated the positive colonies (RBS_B0034-LL37-TT_B0015) and also checked by digestion. The construct was successfully cloned downstream the inducible promoter T5LacOperator (BBa_K875002). The M15 cells (containing the pREP4 plasmid coding for the Lac repressor) were transformed with the construct described above and seeded in Kan (M15 resistance), Cm (pSB1C3 resistance) in two conditions: with and without IPTG ( the inhibitors of the repressor Lac).

Unfortunately the LL-37 cathelicidin was probably unable to disrupt the cell membrane, as we observed a growth both in induced and not induced culture.

Antibody

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Chassis

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