title: [Construction and expression of eukaryotic expression plasmid pcDNA3.1/hIL-18].
abstract: OBJECTIVE
  To construct an eukaryotic expression plasmid pcDNA3.1/hIL-18 and express it in mammalian cells.
  
  
  METHODS
  cDNA encoding mature hIL-18 was cleavaged by enzyme digestion from mesomeric clone vector pGEM-T/hIL-18 and inserted into an eukaryotic expression plasmid pcDNA3.1 to construct a recombinant expression plasmid pcDNA3.1/hIL-18. Then the constructed plasmid was transfected into COS-7 and Rlc310 cells by liposome-mediated gene transfer method. hIL-18 expressed in transfected COS-7 and Rlc310 cells was detected by immunohistochemical staining and level of hIL-18 mRNA in transfected Rlc310 cells was assayed by RT-PCR.
  
  
  RESULTS
  A recombinant eukaryotic expression plasmid pcDNA3.1/hIL-18 was successfully constructed and expressed transiently in COS-7 cells and stably in Rlc310 cells.
  
  
  CONCLUSIONS
  The construction and expression of pcDNA3.1/hIL-18 have been achieved successfully, which lays a foundation for further research on anti-tumor effect of IL-18.
authors.affiliation: Institute for Cancer Research, School of Life Science and Technology, Xi'an Jiaotong University, Xi'an 710061,China.
authors.forename: Ling-Hong
authors.initials: LH
authors.lastname: Kong
authors.forename: Yao-Chen
authors.initials: YC
authors.lastname: Li
authors.forename: Yi-Li
authors.initials: YL
authors.lastname: Wang
authors.forename: Ning-Kuan
authors.initials: NK
authors.lastname: Shang
authors.forename: Bao-Chang
authors.initials: BC
authors.lastname: Lai
authors.forename: Yi-Ping
authors.initials: YP
authors.lastname: Geng
authors.forename: Lu-Sheng
authors.initials: LS
authors.lastname: Si
mesh_terms: D000818:Animals; D019556:COS Cells; D056890:Eukaryota / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors; D006801:Humans; D010957:Plasmids; D014162:Transfection