title: [Construction and identification of rhesus monkey Schwann cells modified with human glial cell derived neurotrophic factor gene].
abstract: OBJECTIVE
  To construct the rhesus monkey Schwann cells (SCs) modified with human glial cell derived neurotrophic factor (hGDNF) gene.
  
  
  METHODS
  The coding sequence ofhGDNF amplified by PCR from pUC19-hGDNF was inserted into eukaryotic expression vector pBABE-puro. The recombinant eukaryotic expression vector pBABE-puro-hGDNF was identified with restriction enzyme digestion and DNA sequencing. The SCs were isolated from rhesus monkeys, cultured and purified. The SCs were transfected with the recombinant retrovirus vector containing hGDNF gene. The mRNA and protein expressions of hGDNF were analyzed by real-time fluorescent quantitative PCR and Western blot.
  
  
  RESULTS
  The PCR product of hGDNF coding sequence was a 596 bp specific segment. The recombinant eukaryotic expression vector was digested into a 596 bp specific segment by specific restriction enzyme and another segment. The 596 bp segment confirmed by DNA sequencing was consistent with hGDNF sequence on GenBank. Restriction enzyme digestion and sequencing results showed that the coding sequence of hGDNF was successfully inserted into the recombinant retrovirus vector and the mRNA and protein expressions of hGDNF were significantly higher in transfected SCs than non-transfected SCs (P < 0.05).
  
  
  CONCLUSIONS
  The rhesus monkey SCs modified with hGDNF gene are successfully constructed and hGDNF can be released continuously and stably, which will provide a foundation for the further research about cell therapy of hGDNF-SCs in the repair of injured nerve.
authors: Department of Neurosurgery, the Third Affiliated Hospital, Sun Yat-sen University, Guangzhou Guangdong, 510630, P.R.China.
authors: Yueyang
authors: Y
authors: Ba
authors: Department of Neurosurgery, the Third Affiliated Hospital, Sun Yat-sen University, Guangzhou Guangdong, 510630, P.R.China.
authors: Hui
authors: H
authors: Wang
authors: Department of Neurosurgery, the Third Affiliated Hospital, Sun Yat-sen University, Guangzhou Guangdong, 510630, P.R.China.
authors: Xinjie
authors: X
authors: Ning
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics*; D051100:Glial Cell Line-Derived Neurotrophic Factor / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D008253:Macaca mulatta; D008297:Male; D010957:Plasmids / Q000235:genetics; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D012190:Retroviridae / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012583:Schwann Cells / Q000378:metabolism*; D014162:Transfection
chemical_list: C495039:GDNF protein, human; D051100:Glial Cell Line-Derived Neurotrophic Factor; D012333:RNA, Messenger; D011994:Recombinant Proteins