Team:USP-UNESP-Brazil/Plasmid Plug n Play/Background

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{{:Team:USP-UNESP-Brazil/Templates/RImage | image=fig3plugplay.JPG | caption= Fig.3. ORF circularization and insertion in Plug&Play plasmid | size=600px }}
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{{:Team:USP-UNESP-Brazil/Templates/RImage | image=Pplay_fig.jpeg | caption= Fig.3. ORF circularization and insertion in Plug&Play plasmid | size=600px }}
This project aims to accomplish the expression of any gene in a two-step process: a PCR reaction and a bacterial transformation. The linear DNA from the PCR is directly inserted into electrocompetent ''E. coli'' cells (e.g. BL21(DE3)) already harboring the Plug&play plasmid and expressing Cre recombinase for the circularization and the integration of the target gene. Mathematical modeling performed to predict rates of linear DNA degradation, integration and circularization showed the viability of the project even before beginning to work in the bench.
This project aims to accomplish the expression of any gene in a two-step process: a PCR reaction and a bacterial transformation. The linear DNA from the PCR is directly inserted into electrocompetent ''E. coli'' cells (e.g. BL21(DE3)) already harboring the Plug&play plasmid and expressing Cre recombinase for the circularization and the integration of the target gene. Mathematical modeling performed to predict rates of linear DNA degradation, integration and circularization showed the viability of the project even before beginning to work in the bench.

Revision as of 11:40, 25 September 2012