title: [Rescue and Amplification of Recombinant Human Adenovirus Type 41 in 293 Cells].
abstract: Human adenovirus type 41 (HAdV-41) is considered to be a "fastidious adenovirus". E1-deleted HAdV-41 cannot be rescued or amplified in 293 cells. To propagate recombinant HAdV-41 in 293 cells, the backbone plasmid pAdbone41 was reconstructed. That is, the E3 coding sequence of HAdV-41 was deleted and replaced with the HAdV-5 E4orf6 gene; and the E1A enhancer of HAdV-5 was inserted upstream of the E4 promoter of HAdV-41. Novel adenoviral plasmid pAd41E4EE-GFP was generated by homologous recombination of the shuttle plasmid pSh41-GFP with the modified backbone plasmid in the Escherichia coli BJ5183 strain. Adenovirus HAdV-41-E4EE-GFP was rescued by transfecting 293 cells with linearized pAd41E4EE-GFP. After seven rounds of propagation, viruses were purified by the CsCl ultracentrifugation method. HAdV-41-E4EE-GFP in 1.0 ml with a particle titer of 8 x 10(10) vp/mL was obtained which had a particle-to-infectious ratio of 50 : 1. The genome of HAdV-41-E4EE-GFP was confirmed by restriction analyses and polymerase chain reaction. These results showed that a novel HAdV-41 vector system was established in which recombinant HAdV-41 could be constructed and packaged in 293 cells.
authors.forename: Xiaohui
authors.initials: X
authors.lastname: Zou
authors.forename: Xiaojuan
authors.initials: X
authors.lastname: Guo
authors.forename: Rong
authors.initials: R
authors.lastname: Xiao
authors.forename: Min
authors.initials: M
authors.lastname: Wang
authors.forename: Zhuozhuang
authors.initials: Z
authors.lastname: Lu
authors.forename: Tao
authors.initials: T
authors.lastname: Hong
mesh_terms: D000260:Adenoviruses, Human / Q000235:genetics* / Q000254:growth & development / Q000502:physiology; D002460:Cell Line; D005822:Genetic Vectors / Q000235:genetics / Q000502:physiology; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D011995:Recombination, Genetic; D014162:Transfection; D019065:Virus Assembly
chemical_list: D049452:Green Fluorescent Proteins