Part:BBa_K320004:Experience

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Applications of BBa_K320004

This BioBrick has two main functions, it makes the bacteria to resistant to the Antibiotic Kanamycin and it provides an origin of replication working for E. coli and Asaia.

To test the BioBrick, we added it in the pSBC3 vector. This plasmid provides a resistance to Chloramphenicole and an origin of replication which works only for E.Coli.

We transfected both E. coli and Asaia with the plasmid.

The transfected E. coli were grown on a plate of LB with Agar containing Kanamycin. This experiment confirmed that the part provides the bacteria with resistance to Kanamycin.

The transfected Asaia were grown on a plate of Gly with Agar containing Kanamycin with a concetration of 100 ng/microl ([http://2010.igem.org/Team:EPF_Lausanne/Project_asaia "characterisation of Asaia"]). This experiment confirmed that the origin of replication worked in Asaia and that the BioBrick conferred resistance to kanamycine.

Of course we also made two negative controls with E. coli or Asaia without plasmid, for which we observed no colonies on any plate.

We conclude that the resistance to Kanamycin and the origin of replication work for both E. coli and Asaia.

User Reviews

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