Rescue of avian adeno-associated virus from a recombinant plasmid containing deletions in the viral inverted terminal repeats. 2012

Jianye Wang, and Liqian Zhu, and Jun Zhu, and Xinjun Zhang, and Jie Tao, and Qiangde Duan, and Guoqiang Zhu
Ministry of Education Key Lab for Avian Preventive Medicine, College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China. wangjy@yzu.edu.cn

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.

UI MeSH Term Description Entries
D008969 Molecular Sequence Data Descriptions of specific amino acid, carbohydrate, or nucleotide sequences which have appeared in the published literature and/or are deposited in and maintained by databanks such as GENBANK, European Molecular Biology Laboratory (EMBL), National Biomedical Research Foundation (NBRF), or other sequence repositories. Sequence Data, Molecular,Molecular Sequencing Data,Data, Molecular Sequence,Data, Molecular Sequencing,Sequencing Data, Molecular
D009690 Nucleic Acid Conformation The spatial arrangement of the atoms of a nucleic acid or polynucleotide that results in its characteristic 3-dimensional shape. DNA Conformation,RNA Conformation,Conformation, DNA,Conformation, Nucleic Acid,Conformation, RNA,Conformations, DNA,Conformations, Nucleic Acid,Conformations, RNA,DNA Conformations,Nucleic Acid Conformations,RNA Conformations
D010322 Parvoviridae Infections Virus infections caused by the PARVOVIRIDAE. Parvovirus Infections,Infections, Parvoviridae,Infections, Parvovirus,Infection, Parvoviridae,Infection, Parvovirus,Parvoviridae Infection,Parvovirus Infection
D010957 Plasmids Extrachromosomal, usually CIRCULAR DNA molecules that are self-replicating and transferable from one organism to another. They are found in a variety of bacterial, archaeal, fungal, algal, and plant species. They are used in GENETIC ENGINEERING as CLONING VECTORS. Episomes,Episome,Plasmid
D002460 Cell Line Established cell cultures that have the potential to propagate indefinitely. Cell Lines,Line, Cell,Lines, Cell
D004279 DNA, Viral Deoxyribonucleic acid that makes up the genetic material of viruses. Viral DNA
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D000229 Dependovirus A genus of the family PARVOVIRIDAE, subfamily PARVOVIRINAE, which are dependent on a coinfection with helper adenoviruses or herpesviruses for their efficient replication. The type species is Adeno-associated virus 2. Adeno-Associated Viruses,Dependoparvovirus,Adeno-Associated Virus,Virus, Adeno-Associated,Viruses, Adeno-Associated,Adeno Associated Virus,Adeno Associated Viruses,Dependoparvoviruses,Dependoviruses,Virus, Adeno Associated,Viruses, Adeno Associated
D001483 Base Sequence The sequence of PURINES and PYRIMIDINES in nucleic acids and polynucleotides. It is also called nucleotide sequence. DNA Sequence,Nucleotide Sequence,RNA Sequence,DNA Sequences,Base Sequences,Nucleotide Sequences,RNA Sequences,Sequence, Base,Sequence, DNA,Sequence, Nucleotide,Sequence, RNA,Sequences, Base,Sequences, DNA,Sequences, Nucleotide,Sequences, RNA
D014779 Virus Replication The process of intracellular viral multiplication, consisting of the synthesis of PROTEINS; NUCLEIC ACIDS; and sometimes LIPIDS, and their assembly into a new infectious particle. Viral Replication,Replication, Viral,Replication, Virus,Replications, Viral,Replications, Virus,Viral Replications,Virus Replications

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