title: [Construction of adeno-associated virus vector containing human papilloma virus 16 E7 gene and its expression in eukaryotic cells].
abstract: OBJECTIVE
  To construct recombinant adeno-associated virus vector containing human papilloma virus (HPV) 16E7 gene and identify its effectiveness of expression in eukaryotic cell.
  
  
  METHODS
  Recombinant adeno-associated virus particles containing HPV16E7 gene were generated by co-transfection of plasmids pAAV-MCS-E7, pAAV-RC and pAAV-helper into HEK293 cells. The viral particles were collected and purified. The vector titer was measured by southern blot. After the eukaryotic cells were transfected by virus particles, RT-PCR and western blot analysis were performed to identify rAAV expression.
  
  
  RESULTS
  The recombinant adeno-associated virus vector containing HPV16E7 gene was correctly constructed. The vector titer measured by southern blot was approximately 1 x 10(11)/ml. After the eukaryotic cells were transfected by the recombinant adeno-associated virus vector, the expression of HPV16E7 gene was identified by RT-PCR and western blot.
  
  
  CONCLUSIONS
  Recombinant adeno-associated virus HPV16E7 vector is successfully constructed and can stably express in eukaryotic cells.
authors.affiliation: Department of Gynecology, Second Affiliated Hospital, Sun Yat-sen University, Guangzhou 510120, China.
authors.forename: Juan
authors.initials: J
authors.lastname: Zhang
authors.forename: Zhong-qiu
authors.initials: ZQ
authors.lastname: Lin
authors.forename: Yong-pai
authors.initials: YP
authors.lastname: Peng
authors.forename: Bing-zhong
authors.initials: BZ
authors.lastname: Zhang
authors.forename: Hui
authors.initials: H
authors.lastname: Zhou
authors.forename: Qing-yuan
authors.initials: QY
authors.lastname: Yang
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D002478:Cells, Cultured; D005260:Female; D005822:Genetic Vectors; D052162:Human papillomavirus 16 / Q000235:genetics*; D006801:Humans; D007668:Kidney / Q000166:cytology; D050725:Papillomavirus E7 Proteins / Q000235:genetics* / Q000378:metabolism
chemical_list: D050725:Papillomavirus E7 Proteins; C059731:oncogene protein E7, Human papillomavirus type 16