title: Construction of an infectious clone of human adenovirus type 41.
abstract: Human adenovirus type 41 (HAdV-41) is well known for its fastidiousness in cell culture. To construct an infectious clone of HAdV-41, a DNA fragment containing the left and right ends of HAdV-41 as well as a kanamycin resistance gene and a pBR322 replication origin was excised from the previously constructed plasmid pAd41-GFP. Using homologous recombination, the plasmid pKAd41 was generated by co-transformation of the E. coli BJ5183 strain with this fragment and HAdV-41 genomic DNA. Virus was rescued from pKAd41-transfected 293TE7 cells, a HAdV-41 E1B55K-expressing cell line. The genomic integrity of the rescued virus was verified by restriction analysis and sequencing. Two fibers on the virion were confirmed by western blot. Immunofluorescence showed that more expression of the hexon protein could be found in 293TE7 cells than in 293 cells after HAdV-41 infection. The feature of non-lytic replication was preserved in 293TE7 cells, since very few progeny HAdV-41 viruses were released to the culture medium. These results show that pKAd41 is an effective infectious clone and suggest that the combination of pKAd41 and 293TE7 cells is an ideal system for virological study of HAdV-41.
authors.affiliation: National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, 100 Ying Xin Jie, Xuan Wu Qu, Beijing 100052, People's Republic of China.
authors.forename: Duo-Ling
authors.initials: DL
authors.lastname: Chen
authors.forename: Liu-Xin
authors.initials: LX
authors.lastname: Dong
authors.forename: Meng
authors.initials: M
authors.lastname: Li
authors.forename: Xiao-Juan
authors.initials: XJ
authors.lastname: Guo
authors.forename: Min
authors.initials: M
authors.lastname: Wang
authors.forename: Xin-Feng
authors.initials: XF
authors.lastname: Liu
authors.forename: Zhuo-Zhuang
authors.initials: ZZ
authors.lastname: Lu
authors.forename: Tao
authors.initials: T
authors.lastname: Hung
mesh_terms: D000260:Adenoviruses, Human / Q000145:classification* / Q000235:genetics* / Q000472:pathogenicity; D000890:Anti-Infective Agents / Q000494:pharmacology; D003001:Cloning, Molecular; D004279:DNA, Viral / Q000235:genetics*; D024882:Drug Resistance, Viral / Q000235:genetics; D016679:Genome, Viral; D057809:HEK293 Cells; D006801:Humans; D007612:Kanamycin / Q000494:pharmacology; D010957:Plasmids; D014779:Virus Replication / Q000502:physiology
chemical_list: D000890:Anti-Infective Agents; D004279:DNA, Viral; D007612:Kanamycin