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2005 | 52 | 3 | 699-702
Article title

Virus-like particles of potato leafroll virus as potential carrier system for nucleic acids.

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EN
Abstracts
EN
Potato leafroll virus is a member of the polerovirus genus. The isometric virion is formed by a coat protein encapsidating single-stranded, positive-sense, mono-partite genomic RNA with covalently attached viral protein at the 5' end. The coat protein of the virus exists in two forms: i) a 23 kDa protein, the product of the coat protein gene, and ii) a 78 kDa protein, the product of the coat protein gene and an additional open reading frame expressed by read-through of the coat protein gene stop codon. The aim of this work was the expression of potato leafroll virus coat protein-based proteins that would be able to assemble into virus-like particles in insect cells. These modified particles were tested for their ability to encapsidate nucleic acids. Two types of N-terminally His-tagged coat protein constructs were used for the expression in insect cells: one, encoding a 23 kDa protein with the C-terminal amino-acid sequence corresponding to the wild type coat protein and the second with additional clathrin binding domain at the C-terminus. The expression of these two proteins by a recombinant baculovirus was characterized by Western immunoblotting with antibodies directed against potato leafroll virus. The protection or putative encapsidation of nucleic acids by these two coat protein derivatives was shown by DNase I and RNase A protection assays.
Year
Volume
52
Issue
3
Pages
699-702
Physical description
Dates
published
2005
received
2005-03-15
revised
2005-07-14
accepted
2005-07-28
References
  • Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith J, Struhl K (1987) Western Blotting. In Current Protocols in Molecular Biology. Ausubel FM, ed, pp 10.8.1-10.8.6. Green Publishing Associates and Wiley-Interscience, New York.
  • Bahner I, Lamb J, Mayo MA, Hay RT (1990) Expression of the genome of potato leafroll virus: readthrough of the coat protein termination codon in vivo. J Gen Virol 71: 2251-2256.
  • Lamb JW, Duncan GH, Reavy B, Gildow FE, Mayo MA, Hay RT (1996) Assembly of virus-like particles in insect cells infected with a baculovirus containing a modified coat protein gene of potato leafroll luteovirus. J Gen Virol 77: 1349-1358.
  • Mayo MA, Robinson DJ, Jolly CA, Hyman L (1989) Nucleotide sequence of potato leafroll luteovirus RNA. J Gen Virol 70: 1037-1051.
  • Palucha A, Sadowy E, Kujawa A, Juszczuk M, Zagorski W, Hulanicka D (1994) Nucleotide sequence of RNA of a Polish isolate of potato leafroll luteovirus. Acta Biochim Polon 41: 405-414.
  • van der Wilk F, Huisman MJ, Cornelissen BJ, Huttinga H, Goldbach R (1989) Nucleotide sequence and organization of potato leafroll virus genomic RNA. FEBS Lett 245: 51-66.
Document Type
Publication order reference
YADDA identifier
bwmeta1.element.bwnjournal-article-abpv52i3p699kz
Identifiers
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