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Applied Microbiology and Biotechnology

, Volume 56, Issue 1–2, pp 150–156 | Cite as

Efficiency of delivery of DNA to cells by bovine papillomavirus type-1 L1/L2 pseudovirions

  •  Y. Liu
  •  I. Frazer
  •  W. Liu
  •  X. Liu
  •  N. McMillan
  •  K.-N. Zhao
Original Paper

Abstract.

To investigate the efficiency of encapsidation of plasmid by papillomavirus virus-like particles (PV VLPs), and the infectivity of the resultant PV pseudovirions, Cos-1 cells were transfected with an 8-kb plasmid incorporating a green fluorescent protein (GFP) reporter gene (pGSV), and infected with bovine PV (BPV-1) L1/L2 recombinant vaccinia virus to produce BPV1 pseudovirions. Approximately 1 in 1.5×104 of dense (1.35 g/ml) PV pseudovirions and 0.3 in 104 of less-dense (1.29 g/ml) pseudovirions packaged an intact pGSV plasmid. The majority (>75%) of packaged plasmids contained deletions, and the deletions affected all tested genes. After exposure of Cos-1 cells to BPV-1 pseudovirions at an MOI of 40,000:1, 6% of cells expressed GFP, giving a calculated efficiency of delivery of the pGSV plasmid, by pseudovirions which had packaged an intact plasmid, of approximately 5%. Plasmid delivery was not effected by purified pGSV plasmid, was blocked by antiserum against BPV-1, and was not blocked by DNase treatment of pseudovirions, confirming that delivery was mediated by DNA within the pseudovirion. We conclude that a major limitation to the use of PV pseudovirions as a gene delivery system is that intact plasmid DNA is not efficiently selected for packaging by VLPs in cell-based pseudovirions production systems.

Keywords

Green Fluorescent Protein Reporter Gene Gene Delivery Vaccinia Virus Vaccinia 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Springer-Verlag 2001

Authors and Affiliations

  •  Y. Liu
    • 1
  •  I. Frazer
    • 1
  •  W. Liu
    • 1
  •  X. Liu
    • 1
  •  N. McMillan
    • 1
  •  K.-N. Zhao
    • 1
  1. 1.Center for Immunology and Cancer Research, University of Queensland, Princess Alexandra Hospital, Woolloongabba, Queensland 4102, Australia

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