title: [Construction of recombinant plasmid containing mouse vasoactive intestinal peptide gene and its expression in COS-7 cells].
abstract: To construct the eukaryotic expression plasmid containing mouse vasoactive intestinal peptide(VIP) gene with biological activities.
  
  
  
  VIP cDNA including the sequences of signal peptide was cloned from mouse thymus by RT-PCR, and then inserted into the mammalian expression vector pcDNA3.1 between Hind III and EcoR I restriction sites. COS-7 cells were transfected with pcDNA3. 1-VIP using liposome, the expression of VIP was identified by Western blot and ELISA. Supernatant of transfected cell culture was added to LPS-stimulated macrophages and the TNF-alpha production in cell medium was observed by ELISA.
  
  
  
  The cloned VIP cDNA was confirmed by enzyme digestion and DNA sequencing. The expression of VIP was detected in the pcDNA3. 1-VIP transfected COS-7 cells by Western blot and ELISA. The VIP in culture supernatant potently inhibited TNF-alpha production by LPS-induced Macrophages in vitro.
  
  
  
  The eukaryotic expression plasmid that expresses biological active murine VIP has been constructed successfully.
authors.affiliation: Institutes of Immunoology, College of Medicine, Zhejiang University, Hangzhou 310031, China.
authors.forename: Song-zhao
authors.initials: SZ
authors.lastname: Zhang
authors.forename: Jian-gen
authors.initials: JG
authors.lastname: Shen
authors.forename: Zhi-xiong
authors.initials: ZX
authors.lastname: Liao
authors.forename: Jian-ping
authors.initials: JP
authors.lastname: Pan
mesh_terms: D000818:Animals; D001483:Base Sequence; D019556:COS Cells; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D005057:Eukaryotic Cells / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D010957:Plasmids; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D011995:Recombination, Genetic; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014660:Vasoactive Intestinal Peptide / Q000096:biosynthesis / Q000235:genetics*
chemical_list: D018076:DNA, Complementary; D011994:Recombinant Proteins; D014660:Vasoactive Intestinal Peptide