Expression of fusion protein of parathyroid hormone and transferrin N-terminal half-molecule in Pichia pastoris / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 804-808, 2005.
Article
Dans Chinois
| WPRIM
| ID: wpr-237070
ABSTRACT
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 Fe3+ in the Fe3+ saturation study as the recombinant TFN did indicating that TFN could be used as the transcellar carrier of PTH.
Texte intégral:
Disponible
Indice:
WPRIM (Pacifique occidental)
Sujet Principal:
Hormone parathyroïdienne
/
Pichia
/
Protéines de fusion recombinantes
/
Transferrine
/
Clonage moléculaire
/
Fusion artificielle de gènes
/
Génétique
/
Métabolisme
Limites du sujet:
Humains
langue:
Chinois
Texte intégral:
Chinese Journal of Biotechnology
Année:
2005
Type:
Article
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