title: [Construction of recombinant vector pEGFP-ANG and expression of angiogenin gene in HUVECs].
abstract: OBJECTIVE
  To express the recombinant angiogenin protein in mammalian cells.
  
  
  METHODS
  Human ANG full-length cDNA was obtained by chemical synthesis. The target gene ANG was inserted into eukaryotic expression vector pEGFP-C2. The recombinant plasmid was transfected into the human umbilical vein endothelial cells (HUVECs) via Lipofectin transfection. The expression of ANG gene in transfected HUVECs was detected by fluorescence microscopy and immunohistochemical staining.
  
  
  RESULTS
  DNA sequencing showed the sequence of the synthetic ANG was correct and amino acid sequence of ANG was right. The green fluorescence could be seen in transfected HUVECs under fluorescence microscope. Immunohistochemical staining detection showed that ANG expressed in transfected cells.
  
  
  CONCLUSIONS
  Human ANG full-length cDNA has been obtained. The ANG protein was expressed in mammalian cells successfully.
authors.affiliation: Department of Medical Research, General Hospital of Guangzhou Military Command, Guangzhou 510010, China. zhangwater@163.net
authors.forename: Hong-bin
authors.initials: HB
authors.lastname: Zhang
authors.forename: Jie
authors.initials: J
authors.lastname: Wang
authors.forename: Jie
authors.initials: J
authors.lastname: Wu
authors.forename: Yue-hua
authors.initials: YH
authors.lastname: Jiang
authors.forename: Tai-cheng
authors.initials: TC
authors.lastname: Yang
authors.forename: Jiang
authors.initials: J
authors.lastname: Xian
authors.forename: Chuan-hong
authors.initials: CH
authors.lastname: Yang
authors.forename: Wen-lin
authors.initials: WL
authors.lastname: Zheng
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D018076:DNA, Complementary / Q000737:chemistry; D042783:Endothelial Cells / Q000378:metabolism*; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis*; D012259:Ribonuclease, Pancreatic / Q000235:genetics*; D014162:Transfection; D014471:Umbilical Veins / Q000166:cytology
chemical_list: D018076:DNA, Complementary; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; C046366:angiogenin; D012259:Ribonuclease, Pancreatic