title: Expression of Recombinant Human Amelogenin in Iranian Lizard Leishmania and Its Biological Function Assay.
abstract: BACKGROUND
  Amelogenins are the major components of enamel matrix proteins. Enamel matrix derivatives (EMD) can be used in periodontal diseases to regenerate periodontal tissues. The main aim of this study was to evaluate expression of full-length functional recombinant human amelogenin (rhAm) in Iranian lizard Leishmania (I.L.L.) as an alternative eukaryotic expression system.
  
  
  METHODS
  Human cDNA encoding a 175-amino acid amelogenin expression cassette was sub cloned into a pLEXSY vector. The construct was transferred into Leishmania cells by electroporation. The protein production was surveyed in the transcription and the translation levels. The expressed protein was purified and some of its biological properties were investigated in comparison to EMD and negative control.
  
  
  RESULTS
  Expression of rhAm was confirmed by RT-PCR and western blot test in Leishmania cells. Purified rhAm significantly inhibited the formation of tartrate-resistant acid phosphatase positive (TRAP(+)) multinuclear cells in calcitriol stimulated mouse marrow cultures. Moreover, it significantly promoted proliferation and DNA synthesis in L929 mouse fibroblast cells.
  
  
  CONCLUSIONS
  Functional rhAm was successfully expressed in I.L.L. Easy handling and post translation modification were the main advantages of this expression system. It is suggested to investigate molecular properties of this rhAm in the future.
authors.affiliation: Dept. of Biotechnology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran ; Dept. of Dental Materials, School of Dentistry, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
authors.forename: Zahra
authors.initials: Z
authors.lastname: Yadegari
authors.affiliation: Dept. of Biotechnology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran ; Dept. of Biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran ; Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
authors.forename: Mojgan
authors.initials: M
authors.lastname: Bandehpour
authors.affiliation: Dept. of Biotechnology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran ; Dept. of Biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran ; Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
authors.forename: Bahram
authors.initials: B
authors.lastname: Kazemi
authors.affiliation: Infectious and Tropical Diseases Research Center, Hormozgan University of Medical Sciences, Bandar Abbas, Iran.
authors.forename: Khojasteh
authors.initials: K
authors.lastname: Sharifi-Sarasiabi
keywords: Amelogenin; Eukaryotic expression system; Iran; Leishmania; Lizard; Osteoclastogenesis; Recombinant protein