title: Construction and identification of polycistron adenoviral expression vector PCA13/FasL-IRES-iNOS.
abstract: OBJECTIVE
  To construct the Fas ligand (FasL) and inducible nitric oxide synthase (iNOS) coexpressed PCA13 plasmid used for packing of adenovirus, so as to observe the immunoprotective effect of FasL on the adenovirus vector.
  
  
  METHODS
  By way of the internal ribosome entry site (IRES), gene engineering techniques such as preparation and transformation of competent cells, plasmid extraction, agarose gel electrophoresis and restriction enzymolysis were used for the construction of the polycistron adenoviral expression vector PCA13/FasL-IRES-iNOS, which could coexpress FasL and the iNOS gene after multisubcloning steps.
  
  
  RESULTS
  FasL and iNOS connected with the IRES were successfully cloned to the PCA13 plasmid and verified by enzymolysis (600 bp FasL, 1000 bp IRES and 4000 bp iNOS) and the gene sequence was concordant with the gene bank.
  
  
  CONCLUSIONS
  The polycistron adenoviral expression vector PCA13/FasL-IRES-iNOS was successfully constructed.
authors.affiliation: Department of Surgery, Renji Hospital, Shanghai Second Medical University, Shanghai, China. zzq72@sina.com.cn
authors.forename: Jiang Feng
authors.initials: JF
authors.lastname: Qiu
authors.forename: Zhi Yong
authors.initials: ZY
authors.lastname: Wu
authors.forename: Chun Xia
authors.initials: CX
authors.lastname: Luo
authors.forename: Fu Rong
authors.initials: FR
authors.lastname: Yu
authors.forename: Zhi Qi
authors.initials: ZQ
authors.lastname: Zhang
mesh_terms: D000256:Adenoviridae / Q000235:genetics* / Q000276:immunology; D053222:Fas Ligand Protein; D018014:Gene Transfer Techniques; D005814:Genes, Viral / Q000235:genetics*; D015316:Genetic Therapy; D005822:Genetic Vectors; D006973:Hypertension / Q000628:therapy; D008562:Membrane Glycoproteins / Q000235:genetics* / Q000276:immunology; D019001:Nitric Oxide Synthase / Q000235:genetics*; D052247:Nitric Oxide Synthase Type II; D010957:Plasmids / Q000235:genetics*; D012270:Ribosomes / Q000378:metabolism; D015678:Viral Structural Proteins / Q000235:genetics*
chemical_list: D053222:Fas Ligand Protein; D008562:Membrane Glycoproteins; D015678:Viral Structural Proteins; D019001:Nitric Oxide Synthase; D052247:Nitric Oxide Synthase Type II