[Construction of C3b(AN42) and OVA(257-264) fusion gene and functional analysis of the fusion protein].

AIM To explore the role of complement system in the T cell activation. METHODS OVA(257-264) CTL epitope and C3b(AN42) encoding genes were synthesized and cloned into expression vector pEGFP-N1. COS-7 cells were transfected with the recombinant plasmid pEGFP-N1. The expression of the fusion protein was determined by FACS. Twenty-four hours after transfection, cultural supernatants of the transfected cells were collected and added to Ana-1 cell culture. Laser confocal microscopy was used to detect the binding and presentation of the fusion protein by Ana-1 cells. RESULTS C3b(AN42)-OVA(257-264) fusion protein was expressed in COS-7 cells. The fusion protein could bind to Ana-1 cells and OVA(257-264) CTL epitope was cross-presented on the surface of Ana-1 cells. CONCLUSION The C3b(AN42)-OVA(257-264) fusion protein can express in COS-7 cells and possesses natural biological activity.