The MGC platform has been created and is managed by both IGMM and CRBM.
It is affiliated to the BioCampus UMS. Its Operational Head is Frédéric Lionneton (CNRS IR, BioCampus) and its staff members are Sylvie Fromont (CNRS AI, CRBM) and Marie-Cécile Robert (CNRS IE part time, IGMM). Its Scientific Head is Edouard Bertrand (CNRS DR1), group leader at IGMM.
The MGC platform manages 2 human ORFeome libraries. The version 5.1 which represents 15483 human ORF clones corresponding to 12794 human genes, and the version 8, which contains 12 230 human clones (11 149 non-redundant ORFs). The clones exclude the 5’ and 3’ untranslated region, and the ORFs are in-frame with the attB recombination sites. The termination codons have also been removed to allow for the addition of a 3’ tag to the ORF of interest. The ORFs have been cloned into the pDONR223 vector (Spectinomycin resistant). To know if your gene of interest is cloned into the hORFome, connect to this website for the version 5.1, or on this website for the version 8 ) and follow the handbook instructions (How to find an ORF clone?).
The Gateway technology is based on site-specific recombination reactions derived from bacteriophage lambda. This method permits the directional recombination of two DNA fragments of carrying Att sites, and it is used here as a high-throughput cloning method. Clones from the ORFeome (Entry vectors) are recombined with a variety of destination vectors carrying a large choice of tags, and adapted to express the ORF in the organism of interest (from E. Coli to mammals). Entry vectors are created by recombining a PCR product that contains the sequence of interest (flanked by two attB recombination sites included into the PCR primers), and a donor vector (pDONR carrying two recombination sites attP).