Labelling of E. coli and human cells with stable isotopes

Labelling of E. coli and human cells with stable isotopes

Description
The main steps for bacterial expression include the construction of expression vectors, the transformation of the bacterial host with these vectors, the expression of the samples in the host, and the purification of the samples.
Several different cloning techniques are currently available; sample expression range from expression of single proteins to the multi-expression of protein complexes. Labelled protein with 15N, 13C and 2H can be obtained.
The strategy for protein expression in mammalian cells is the transfection with plasmid vectors or infection with viral vectors (inducible or constitutive) of cell culture (adherent or suspension). The Instruct center CERM/CIRMMP will perform cell cultures ready for transfection or infection with the vector carrying the recombinant gene provided by the user. In a second step, the Instruct center CERM/CIRMMP will propose a set of expression vectors (commercial or developed on site) to be screened together with the cell types for the optimization of functional and soluble protein and protein complexes production.

Service provider
Consorzio Interuniversitario Risonanze Magnetiche di Metallo Proteine (CIRMMP),
Centro di Risonanze Magnetiche 

Location
Sesto Fiorentino, Italy

Contacts
Scientific contact: Lucia Banci
Technical contact: Rebecca Del Conte