title: Rescue of avian adeno-associated virus from a recombinant plasmid containing deletions in the viral inverted terminal repeats.
abstract: We have previously reported the complete genome sequence of avian adeno-associated virus (AAAV) strain YZ-1, isolated from healthy chickens in China. In this study, we describe the successful rescue of infectious virions from a recombinant plasmid containing the genome of YZ-1 with deletions in the viral inverted terminal repeats (ITRs). The complete genome of YZ-1 was cloned into a bacterial plasmid by a modified "A-T" cloning method. Six recombinant plasmids were selected for further experiments. Sequence analysis indicated that the six clones shared identical internal sequences except for the various deletions within ITRs at either end of the cloned genome. The recombinant plasmid pYZ525, harboring a YZ-1 genome with a 96-nt deletion at the 5' end, was used to transfect CEL or HEK293 cells in the presence of the CELO virus or a helper plasmid, and rescued virions were obtained by both of the methods despite the presence of the deletions. Here, for the first time, we provide evidence that a certain number of nt deletions in the ITRs are not lethal for the rescue of viable AAAV from recombinant plasmids. This study provides insight into the unique biology of AAAV and the mechanism of viral replication.
authors.affiliation: Ministry of Education Key Lab for Avian Preventive Medicine, College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China. wangjy@yzu.edu.cn
authors.forename: Jianye
authors.initials: J
authors.lastname: Wang
authors.forename: Liqian
authors.initials: L
authors.lastname: Zhu
authors.forename: Jun
authors.initials: J
authors.lastname: Zhu
authors.forename: Xinjun
authors.initials: X
authors.lastname: Zhang
authors.forename: Jie
authors.initials: J
authors.lastname: Tao
authors.forename: Qiangde
authors.initials: Q
authors.lastname: Duan
authors.forename: Guoqiang
authors.initials: G
authors.lastname: Zhu
mesh_terms: D001483:Base Sequence; D002460:Cell Line; D004279:DNA, Viral / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D000229:Dependovirus / Q000737:chemistry / Q000235:genetics* / Q000502:physiology; D016679:Genome, Viral; D006801:Humans; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D010322:Parvoviridae Infections / Q000821:virology; D010957:Plasmids / Q000235:genetics* / Q000378:metabolism; D017384:Sequence Deletion; D020079:Terminal Repeat Sequences; D014779:Virus Replication
chemical_list: D004279:DNA, Viral