title: Expression and purification of a recombinant Fip-fve protein from Flammulina velutipes in baculovirus-infected insect cells.
abstract: OBJECTIVE
  To develop an efficient and facile expression system supply of high purity and stable activity of rFip-fve for oral administration, medicinal study and applications.
  
  
  RESULTS
  A recombinant virus that contained the chimera gene, encoding a bombyxin signal peptide sequence fused to a Fip-fve-6His sequence, was constructed. The rFip-fve was purified from the supernatant of the infected Sf21 cells using a nickel-chelated affinity column, and was verified by Western blot and MALDI-MS (matrix-assisted laser desorption ionization mass spectrometry) analyses. Results showed that a glycosylated mature rFip-fve was produced and secreted into the infected cell supernatant. The immunomodulatory activity of rFip-fve was evaluated by measuring the amount of interleukin-2 released from murine splenocytes.
  
  
  CONCLUSIONS
  A reliable scheme to express and purify active rFip-fve in a baculovirus/insect cell system for medicinal applications and genetic study is a feasible means of solving potential problems related to the production and activity of rFip-fve protein.
  
  
  CONCLUSIONS
  The rFip-fve expressed in insect cells was processed and modified in a manner more similar to that of its native counterpart than that in bacterial cells. Therefore, the potential applications of rFip-fve that is generated in Sf21 cells can be more effectively evaluated that produced in Escherichia coli.
authors.affiliation: Biopesticide Department, Taiwan Agricultural Chemicals and Toxic Substances Research Institute, Wufeng, Taiwan.
authors.forename: C-M
authors.initials: CM
authors.lastname: Wu
authors.forename: T-Y
authors.initials: TY
authors.lastname: Wu
authors.forename: S-S
authors.initials: SS
authors.lastname: Kao
authors.forename: J-L
authors.initials: JL
authors.lastname: Ko
authors.forename: T-R
authors.initials: TR
authors.lastname: Jinn
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000502:physiology*; D019149:Bioreactors; D002460:Cell Line; D002478:Cells, Cultured; D004926:Escherichia coli; D005656:Fungal Proteins / Q000302:isolation & purification* / Q000494:pharmacology; D007155:Immunologic Factors / Q000302:isolation & purification* / Q000494:pharmacology; D007218:Industrial Microbiology; D007376:Interleukin-2 / Q000096:biosynthesis; D037102:Lectins / Q000302:isolation & purification* / Q000494:pharmacology; D051379:Mice; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000494:pharmacology; D013154:Spleen / Q000187:drug effects / Q000276:immunology; D018411:Spodoptera / Q000821:virology
chemical_list: D005656:Fungal Proteins; D007155:Immunologic Factors; D007376:Interleukin-2; D037102:Lectins; D011994:Recombinant Proteins