title: [Construction of pORF-mIG and activity evaluation of MIG in vitro].
abstract: OBJECTIVE
  To establish a transformant of monokine induced by interferon-gamma (MIG) with the eukaryotic expression vector for further investigating the efficacy of its use in antitumor gene therapy.
  
  
  METHODS
  The MIG full-length cDNA was amplified from pBLAST-MIG and was cloned into the eukaryote expression vector pORF-mcs, and the resulted recombinant plasmid was named pORF-MIG. The recombinant pORF-MIG was determined with restriction enzyme and sequencing, and then it was transfected into COS-7 cells by Lipfectamin. Expression of the transformant was detected by immunoblot assay, and the bioactivity was assessed by chemotaxis assay.
  
  
  RESULTS
  The restriction analysis and the sequence determination confirmed that the recombinant pORF-MIG contained the MIG full-length cDNA. And the transformants of pORF-MIG expressed the MIG protein which could apparently attract the activated lymphocytes.
  
  
  CONCLUSIONS
  The recombinant pORF-MIG was constructed successfully, and this recombinant eukaryotic expression vector could be used in additional studies on the biological effect of MIG and its use in anti-tumor gene therapy.
authors.affiliation: Laboratory of Biotherapy, Cancer Center, West China Hospital, Sichuan University, Chengdu 610041, China.
authors.forename: Ru
authors.initials: R
authors.lastname: Zhang
authors.forename: Ling
authors.initials: L
authors.lastname: Tian
authors.forename: Fei
authors.initials: F
authors.lastname: Xiao
authors.forename: Yan-jun
authors.initials: YJ
authors.lastname: Wen
authors.forename: Jiong
authors.initials: J
authors.lastname: li
authors.forename: Jian-mei
authors.initials: JM
authors.lastname: Hou
authors.forename: Ling
authors.initials: L
authors.lastname: Zhang
authors.forename: Gang
authors.initials: G
authors.lastname: Li
authors.forename: Bing
authors.initials: B
authors.lastname: Yao
authors.forename: Xian-cheng
authors.initials: XC
authors.lastname: Chen
authors.forename: Kai
authors.initials: K
authors.lastname: Mel
mesh_terms: D001483:Base Sequence; D054370:Chemokine CXCL9; D019743:Chemokines, CXC / Q000096:biosynthesis* / Q000235:genetics; D005057:Eukaryotic Cells / Q000378:metabolism; D015316:Genetic Therapy; D005822:Genetic Vectors; D006801:Humans; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D014162:Transfection
chemical_list: C080810:CXCL9 protein, human; D054370:Chemokine CXCL9; D019743:Chemokines, CXC; D011994:Recombinant Proteins