[Expression of fusion protein of parathyroid hormone and transferrin N-terminal half-molecule in Pichia pastoris]

Sheng Wu Gong Cheng Xue Bao. 2005 Sep;21(5):804-8.
[Article in Chinese]

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.

Publication types

  • English Abstract
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Artificial Gene Fusion
  • Cloning, Molecular
  • Humans
  • Parathyroid Hormone / genetics*
  • Pichia / genetics
  • Pichia / metabolism
  • Recombinant Fusion Proteins / biosynthesis*
  • Recombinant Fusion Proteins / genetics
  • Transferrin / genetics*

Substances

  • N-terminal monoferrictransferrin
  • PTH protein, human
  • Parathyroid Hormone
  • Recombinant Fusion Proteins
  • Transferrin