title: [Study on construction of chimeric adenovirus vector Ad5/11 carrying human eGFP and endostatin-K5 and its experimental investigation in vitro].
abstract: OBJECTIVE
  To construct chimeric adenoviral vector Ad5/11 carrying reporter gene eGFP and human endostatin-K5.
  
  
  METHODS
  Chimeric adenoviral backbone vector expressing eGFP was generated by overlap PCR and homologous recombination in E.coli BJ5183. Then chimeric adenoviral vector Ad5/11-E1-CMV-endo-K5/E3-CMV-eGFP carrying eGFP and human endostatin-K5 was constructed by co-transfecting Pac I linearized chimeric adenoviral backbone and adenoviral E1 shuttle vector expressing human endostatin-K5 into HEK 293 cells. The expression of eGFP was observed under fluorescent microscope. The expression of human endostain-K5 in U87MG cells infected by chimeric adenoviral vector was detected by RT-PCR. The infection efficiency between chimeric adenovirus and unmodified control adenovirus for human glioblastoma cell line A172 and breast cancer cell line MDA-MB-231 in vitro was evaluated by the comparison of the expression of eGFP.
  
  
  RESULTS
  Chimeric adenovirus Ad5/11-E1-CMV-endo-K5/E3-CMV-eGFP could successfully express eGFP and endostatin-K5. Chimeric adenoviral vector significantly enhances the infection efficiency for human glioblastoma cell line A172 and breast cancer cell line MDA-MB-231 compared with unmodified adenoviral vector Ad5 E1-CMV-eGFP.
  
  
  CONCLUSIONS
  Chimeric adenoviral vector Ad5/11-E1-CMV-endo-K5/E3-CMV-eGFP can significantly improve the infection efficiency for human glioblastoma cell line A172 and breast cancer cell line MDA-MB-231.
authors.affiliation: Laboratory of Gene Therapy, College of Life Sciences, Shaanxi Normal University, Xi'an 710062, China. wangdongyang007@yahoo.com.cn
authors.forename: Dong-Yang
authors.initials: DY
authors.lastname: Wang
authors.forename: Shi-Hai
authors.initials: SH
authors.lastname: Liu
authors.forename: Xing
authors.initials: X
authors.lastname: Li
authors.forename: Jun-Li
authors.initials: JL
authors.lastname: Zhao
authors.forename: Hao
authors.initials: H
authors.lastname: Chen
authors.forename: Fei-Xue
authors.initials: FX
authors.lastname: Feng
authors.forename: Li-Xian
authors.initials: LX
authors.lastname: Wang
authors.forename: Hai-Bin
authors.initials: HB
authors.lastname: Xia
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D045744:Cell Line, Tumor; D043169:Endostatins / Q000235:genetics* / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000235:genetics; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000235:genetics* / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans
chemical_list: D043169:Endostatins; D049452:Green Fluorescent Proteins