Suicide System

Finally we got the antimicrobial peptide LL37 in pEX-A plasmid and we amplified it through the transformation of DH5-alpha bacteria.

The positive colonies were then digested EcoRI/SpeI, the fragment was eluted from the gel and then cut it again with EcoRI, because the pEX-A plasmid (link alla foto scannerizata) has another EcoRI restriction site downstream our fragment.

Then we did a column purification of the fragment and we successfully cloned the LL-37 cut EcoRI/PstI into the pSB1C3 linearized backbone to make it ready for a potential submission.

We cut the plasmid with EcoRI/SpeI to insert it into the B0015 previously cut EcoRI/XbaI, we purified and the DH5alpha were transformed with the plasmid (LL-37-B0015). The positive colonies were inoculated and then cloned downstream the RBS_B0034.

We also cloned the T5LacOperator inside the pSB1C3 plasmid, making it ready for the submission.

Antibody

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Chassis

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