GP60 of Cryptosporidium parvum: heterologous expression in Tetrahymena thermophila as vaccine candidate

ELGUERO ME; TOMAZIC ML; MONTES MG; NUDEL CB; SCHNITTGER L; NUSBLAT AD

Congreso; Sociedad Argentina de Investigacion en Bioquimica y Biologia Molecular; 2015

Cryptosporidium parvum is a protozoan parasite of the phylum Apicomplexa which is responsible of cryptosporidiosis, a disease that generates significant economic deficits in livestock. One of the main determinants of the different subtypes of Cryptosporidium is the surface protein GP60. This protein like most phylum surface proteins is attached to the membrane by a glycophosphatidylinositol anchor-type (GPI) and often involved in recognition processes, adhesion and invasion of host cells. These characteristics make the GPI proteins essential for the survival of protozoa and also turn into vaccine candidates in high biotechnological value. In this work the heterologous expression of GP60 antigen from C. parvum was held in the ciliate Tetrahymena thermophila using ribosomal DNA vectors. The presence of GP60 was evaluated by western blot with specific antibodies. The protein is expressed in a 40 kDa variant and is localized in an insoluble fraction after Triton X-114 phase-partitioning procedure. It was also detected by Coomasie Brilliant Blue staining and confirmed by HPLC-MS indicating that this ciliated GPI is not only able to express but also to perform the post-translational modifications necessary for obtaining GP60 antigen from C. parvum.Financed by PICT-2013-1708 y PNSA-1115053 de INTA