title: [Preparation of lyophilized lentiviral vector of bone morphogenetic protein 2].
abstract: OBJECTIVE
  To construct recombinant lentiviral vectors of bone morphogenetic protein 2(BMP-2) gene and prepare a stable lyophilized state.
  
  
  METHODS
  The BMP-2 gene lentiviral vector was constructed by recombinant DNA technology and mixed with different stabilizers in an appropriate proportion and then lyophilized. The optimum stabilizer was selected according to the appearance and virus titer after being lyophilized. The quality of lyophilized product was measured by thermal stability of the virus, PCR and gene sequencing. SPSS13.0 software package was used for statistical analysis.
  
  
  RESULTS
  Recombinant BMP-2 lentiviral vector was successfully constructed. 10% trehalose, 1% bovine serum albumin, 3%mannitol and 0.5% gelatin in group B showed good protection on BMP-2 gene lentiviral vector. The virus titer decreased 0.42 LgPFU/mL after being lyophilized, which was better than group A, C and the control group and the difference was statistically significant (P<0.05); Lyophilized protective agent made from group B still maintained a good appearance at 37 degrees centigrade after 28 days and the virus titer decreased 0.63 LgPFU/mL. The virus titer in liquid lentiviral infection control group decreased rapidly to 2.37 LgPFU/mL 1 week later and the difference between the two groups was statistically significant (P<0.05); PCR and gene sequencing showed that the target gene information after redissolve had no loss or mutation.
  
  
  CONCLUSIONS
  Appropriate selection of lyoprotectants can effectively protect the biological stability of recombinant BMP-2 gene lentiviral vector.
authors.affiliation: Department of Endodontics, Shanghai Stomatological Disease Center. Shanghai 200001, China.E-mail:sammi510@126.com.
authors.forename: Xiao-ling
authors.initials: XL
authors.lastname: Wei
authors.forename: Rong-jing
authors.initials: RJ
authors.lastname: Chen
authors.forename: Jun
authors.initials: J
authors.lastname: Zhao
authors.forename: Zheng
authors.initials: Z
authors.lastname: Hu
mesh_terms: D055396:Bone Morphogenetic Protein 2 / Q000235:genetics*; D005822:Genetic Vectors; D016086:Lentivirus; D011994:Recombinant Proteins / Q000235:genetics; D014162:Transfection; D016212:Transforming Growth Factor beta / Q000235:genetics
chemical_list: D055396:Bone Morphogenetic Protein 2; D011994:Recombinant Proteins; D016212:Transforming Growth Factor beta; C499469:recombinant human bone morphogenetic protein-2