Expression of Fusion Protein of Parathyroid Hormone and Transferrin N-terminal Half-molecule in Pichia pastoris
甲状旁腺激素和转铁蛋白N端半分子融合蛋白在毕赤酵母中的表达

The fused gene (PTH_TFN) of parathyroid hormone (PTH) gene and transferring N_terminal half_molecule (TFN) gene was amplified by multiple PCR and inserted into pPIC9 vector. The recombinant plasmid pPIC9_PTH_TFN was transformed into Pichia pastoris GS115 by PEG. After methanol induction, the target protein was expressed in fermentation supernatant at high level.The fused protein PTH_TFN with purity being higher than 95% was finally obtained after purification through two_step chromatography : SP Sepharose Fast Flow and Phenyl Sepharose Fast Flow.Western blot analysis and adenylate cyclase assay proved that the fused protein exhibited the bioactivity to stimulate cAMP synthesis and the ability to bind Fe ~3+ in the Fe ~3+ saturation study as the recombinant TFN did indicating that TFN could be used as the transcellar carrier of PTH.