title: [Construction and identification of recombinant adenoviruses containing the gene fragments encoding Her2/neu extracellular and transmembrane domains].
abstract: OBJECTIVE
  To construct eukaryotic expression vectors using recombinant adenovirus containing the gene fragments encoding Her2/neu extracellular first ligand-binding domain (Her2-ECD), full-length extracellular domain (Her2-ECD), and extracellular and transmembrane domain (Her2-TM).
  
  
  METHODS
  The cDNAs were amplified by RT-PCR and inserted into shuttle pAdTrack-CMV plasmids. Viral plasmids were obtained from homologous recombination in E. coli BJ5183, and transfected into 293 cells via liposome. Formation of viral plaque and expression of green fluorescent protein were observed by fluorescence microscopy, and the target proteins were detected by Western blotting.
  
  
  RESULTS
  The target cDNA fragments were amplified by PCR with expected lengths and the DNA sequences were confirmed against Genbank. Formation of viral plaque, expression of green fluorescent protein and the target proteins were detected in 293 cells transfected by the viral plasmids, which showed elevated expression of Her2/neu protein with the increase of multiplicity of infection (MOI).
  
  
  CONCLUSIONS
  The eukaryotic expression vectors using recombinant adenovirus have been successfully constructed for expression of Her2/neu extracellular and transmembrane domains.
authors.affiliation: Department of Oncology, Nanfang Hospital, Department of Radiation Medicine, Southern Medical University, Guangzhou 501515, China. mashudong@yahoo.com.cn
authors.forename: Shu-dong
authors.initials: SD
authors.lastname: Ma
authors.forename: Rong-cheng
authors.initials: RC
authors.lastname: Luo
authors.forename: Zheng-hua
authors.initials: ZH
authors.lastname: Ding
authors.forename: Chang-qing
authors.initials: CQ
authors.lastname: Yuan
authors.forename: Xue-mei
authors.initials: XM
authors.lastname: Ding
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D015153:Blotting, Western; D002460:Cell Line; D003001:Cloning, Molecular; D005057:Eukaryotic Cells / Q000166:cytology / Q000378:metabolism; D015870:Gene Expression; D005822:Genetic Vectors / Q000737:chemistry / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008081:Liposomes / Q000737:chemistry; D008856:Microscopy, Fluorescence; D010446:Peptide Fragments / Q000235:genetics / Q000378:metabolism; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection / Q000379:methods
chemical_list: D049452:Green Fluorescent Proteins; D008081:Liposomes; D010446:Peptide Fragments; D018719:Erb-b2 Receptor Tyrosine Kinases; D011993:Recombinant Fusion Proteins