Molecular cloning and characterization of growth factor receptor bound-protein in Clonorchis sinensis

초록

Clonorchis sinensis causes very serious clonorchiasis. Growth factor receptor bound-protein 2 (Grb2) is a very important protein that is located in cytoplasm. Grb2 is found to be conserved in many animals and plays very important roles in many cellular functions and pathological processes including meiosis, organgenosis and even in energy metabolism. It is not know whether there is a Grb2 in C. sinesis to control its development. This study was performed to identify a grow factor receptor bound protein in C. sinesis (CsGFBP) and to characterize its possible functions in organogenesis and development of C. sinensis. A C. sinensis cDNA clone (CsGFBP) encoding putative polypeptide of 311 amino acids with the predicted SH3-SH2-SH3 structure was retrieved from the C. sinensis transcriptome. Recombinant CsGFBP was expressed and purified bacterially. Native CsGFBP was recognized from C. sinensis adult extract by western blotting using a mouse immune serum raised against recombinant CsGFBP. CsGFBP was expressed higher in the metacercariae than in the adult determined by quantitative real time PCR. By immunohistochemical staining with CsGFBP immune mouse sera, CsGFBP was localized in oral sucker, mesenchymal tissues, sperms in seminal receptal and ovary of the adults and abundantly in most organs of the metacercariae. The CsGFBP was highly conserved and suggested to play roles in organogenesis, energy metabolism and sperm production of C. sinensis.