title: [Construction of human bone morphogenetic protein 2 and histidine eukaryotic expression plasmid and synthesis of chitosan/pIRES2-EGFP-hBMP2-His nanoparticles].
abstract: OBJECTIVE
  To clone and construct a eukaryotic expression vector of human bone morphogenetic protein (BMP) 2 and histidine in vitro and synthesize chitosan (CS)/pIRES2-EGFP-hBMP2-His nanoparticles.
  
  
  METHODS
  pMD18T-hBMP2-His was digested by EcoR I and BamH I to obtain the hBMP2-His gene, which was inserted into pIRES2-EGFP to form pIRES2-EGFP-hBMP2-His. Afterward, CS, which exhibited five different molecular weights and deacetylation degrees, was complexed with pIRES2-EGFP-hBMP2-His to form CS/pIRES2-EGFP-hBMP2-His nanoparticles; in this procedure, a desolvent method was used at different N/P ratios (amino in CS to phospho in plasmid DNA). The gene-encapsulating ability of CS was evaluated by agarose gel electrophoresis and fluorescence spectrophotometry; size, distribution, and potential were analyzed using a ZetaPALS analyzer. The shape of the nanoparticles was observed under an atomic force microscope.
  
  
  RESULTS
  1) pIRES2-EGFP-hBMP2-His was constructed after the cloned hBMP2-His gene was confirmed by sequencing. 2) CS/pIRES2-EGFP-hBMP2-His nanoparticles were synthesized and pIRES2-EGFP-hBMP2-His was packaged by CS. 3) CS/pIRES2-EGFP-hBMP2-His nanoparticles were globular with an average size of 111.7 nm to 3,214.2 nm and an average zeta-potential of 4.93 mV to 16.79 mV.
  
  
  CONCLUSIONS
  CS/pIRES2-EGFP-hBMP2-His nanospheres are successfully synthesized.
authors.forename: Yang
authors.initials: Y
authors.lastname: Xiaoyu
authors.forename: Li
authors.initials: L
authors.lastname: Shiyi
authors.forename: Zhang
authors.initials: Z
authors.lastname: Di
authors.forename: Wu
authors.initials: W
authors.lastname: Ying
authors.forename: Yang
authors.initials: Y
authors.lastname: Tao
authors.forename: Liu
authors.initials: L
authors.lastname: Changhong
mesh_terms: D055396:Bone Morphogenetic Protein 2; D048271:Chitosan; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D006639:Histidine; D006801:Humans; D053758:Nanoparticles; D010957:Plasmids
chemical_list: C525718:BMP2 protein, human; D055396:Bone Morphogenetic Protein 2; D049452:Green Fluorescent Proteins; D006639:Histidine; D048271:Chitosan