title: [Construction and identification of a vector inserted with gene of T7 RNA polymerase].
abstract: OBJECTIVE
  To develop a system to rescue virus by intracellular expression of T7 RNA Polymerase.
  
  
  METHODS
  The gene of T7 RNA Polymerase was amplified and cloned to VR1012 by molecular biological technology. The expression plasmid VR-1a was then identified. VR-1a and EV71 infectious plasmid were co-transfected in Vero cell. CPE was observed and viral gene viral antigen were detected.
  
  
  RESULTS
  The gene of T7 RNA Polymerase was successfully cloned into vector VR1012. Vero cell developed to CPE after being transfected VR-1a and EV71 infectious plasmid. EV71 gene was amplified by RT-PCR from the culture. EV71 antigen was also detected by ELISA.
  
  
  CONCLUSIONS
  The method can be used to rescue virus. It could apply to immunologic research of EV71 DNA vaccine.
authors.affiliation: College of Life Sciences, Jilin University, Changchun, 130012, China.
authors.forename: Hong-hui
authors.initials: HH
authors.lastname: Shen
authors.forename: Bing-ke
authors.initials: BK
authors.lastname: Bai
authors.forename: Hao-dong
authors.initials: HD
authors.lastname: Liu
authors.forename: Sheng-dong
authors.initials: SD
authors.lastname: Luo
authors.forename: Yan
authors.initials: Y
authors.lastname: Hu
authors.forename: Jun
authors.initials: J
authors.lastname: Hou
authors.forename: Zhi-jie
authors.initials: ZJ
authors.lastname: Wang
authors.forename: Wei
authors.initials: W
authors.lastname: Kong
authors.forename: Yi-dan
authors.initials: YD
authors.lastname: Bao
authors.forename: Pan-yong
authors.initials: PY
authors.lastname: Mao
mesh_terms: D000818:Animals; D002522:Chlorocebus aethiops; D012321:DNA-Directed RNA Polymerases / Q000235:genetics* / Q000378:metabolism*; D029821:Enterovirus A, Human / Q000235:genetics / Q000502:physiology; D015870:Gene Expression; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics* / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D014162:Transfection; D014709:Vero Cells; D014764:Viral Proteins / Q000235:genetics* / Q000378:metabolism*; D014779:Virus Replication
chemical_list: D014764:Viral Proteins; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases