title: Application of baculovirus as a delivery vehicle for study of transcription and translation mechanism of parvovirus in non-permissive mammalian cells.
abstract: Human bocavirus (HBoV) is a parvovirus associated with respiratory and gastrointestinal diseases in humans. Recombinant baculoviruses have been used widely for many years to transduce transiently and stably a variety of mammalian cell types at high frequencies. In this study, to explore further the use of baculovirus as a gene delivery vehicle for study of transcription and translation mechanism of human bocavirus which lacks susceptible cell culture system, two recombinant baculoviruses were constructed: Bac-BoV-EGFP in which the EGFP gene was under the control of the HBoV1 promoter, and Bac-HBoV1 encompassing the nearly whole HBoV1 genome without both termini. The data demonstrated that efficient gene delivery and expression were observed in numerous mammalian cells transduced by Bac-BoV-EGFP and the transduction rate was much greater than that in plasmid-based transfected cells. The analysis of transcription and translation in Bac-HBoV1 transduced A549 cells showed that two transcripts from NP1 gene were detected by RT-PCR and the NP1 was localized in the nucleus, suggesting that the Bac-HBoV1 recombinant baculovirus delivered efficiently the HBoV1 genome into A549 cells. In summary, this system provides a useful tool for analysis of the transcription and translation of some viruses lacking a virus-cell replication system.
authors.affiliation: College of Life Science, Central China Normal University, Wuhan 430079, Hubei, China.
authors.forename: Jingjing
authors.initials: J
authors.lastname: Li
authors.forename: Bin
authors.initials: B
authors.lastname: Sun
authors.forename: Hu
authors.initials: H
authors.lastname: Han
authors.forename: Jinfeng
authors.initials: J
authors.lastname: Ouyang
authors.forename: Hanchao
authors.initials: H
authors.lastname: Yao
authors.forename: Yongbo
authors.initials: Y
authors.lastname: Yang
authors.forename: Kaiyu
authors.initials: K
authors.lastname: Liu
authors.forename: Yi
authors.initials: Y
authors.lastname: Li
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D002460:Cell Line; D015967:Gene Expression Regulation, Viral; D005822:Genetic Vectors; D056661:Human bocavirus / Q000235:genetics*; D006801:Humans; D011401:Promoter Regions, Genetic; D014176:Protein Biosynthesis; D011995:Recombination, Genetic; D014158:Transcription, Genetic; D014773:Virology / Q000379:methods