title: Human granulocyte colony-stimulating factor (hG-CSF) expression in plastids of Lactuca sativa.
abstract: BACKGROUND
  Human granulocyte colony-stimulating factor (hG-CSF) can serve as valuable biopharmaceutical for research and treatment of the human blood cancer. Transplastomic plants have been emerged as a new and high potential candidate for production of recombinant biopharmaceutical proteins in comparison with transgenic plants due to extremely high level expression, biosafety and many other advantages.
  
  
  METHODS
  hG-CSF gene was cloned into pCL vector between prrn16S promoter and TpsbA terminator. The recombinant vector was coated on nanogold particles and transformed to lettuce chloroplasts through biolistic method. Callogenesis and regeneration of cotyledonary explants were obtained by Murashige and Skoog media containing 6-benzylaminopurine and 1-naphthaleneacetic acid hormones. The presence of hG-CSF gene in plastome was studied with four specific PCR primers and expression by Western immunoblotting.
  
  
  RESULTS
  hG-CSF gene cloning was confirmed by digestion and sequencing. Transplastomic lettuce lines were regenerated and subjected to molecular analysis. The presence of hG-CSF in plastome was confirmed by PCR using specific primers designed from the plastid genome. Western immunoblotting of extracted protein from transplastomic plants showed a 20-kDa band, which verified the expression of recombinant protein in lettuce chloroplasts.
  
  
  CONCLUSIONS
  This study is the first report that successfully express hG-CSF gene in lettuce chloroplast. The lettuce plastome can provide a cheap and safe expression platform for producing valuable biopharmaceuticals for research and treatment.
authors.affiliation: Molecular Systems Biology Group of Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran.
authors.forename: Mehdi
authors.initials: M
authors.lastname: Sharifi Tabar
authors.forename: Ali Akbar
authors.initials: AA
authors.lastname: Habashi
authors.forename: Hamid
authors.initials: H
authors.lastname: Rajabi Memari
mesh_terms: D002736:Chloroplasts / Q000378:metabolism; D003001:Cloning, Molecular; D015870:Gene Expression; D005822:Genetic Vectors; D018745:Genome, Plant / Q000235:genetics; D005966:Glucuronidase / Q000378:metabolism; D016179:Granulocyte Colony-Stimulating Factor / Q000235:genetics*; D006801:Humans; D015151:Immunoblotting; D018545:Lactuca / Q000235:genetics*; D030821:Plants, Genetically Modified; D018087:Plastids / Q000235:genetics*; D016133:Polymerase Chain Reaction; D012038:Regeneration
chemical_list: D016179:Granulocyte Colony-Stimulating Factor; D005966:Glucuronidase