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Stop codon insertio...
Stop codon insertion restores the particle formation ability of hepatitis B virus core-hantavirus nucleocapsid protein fusions
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Kazaks, A (författare)
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Lachmann, S (författare)
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Koletzki, D (författare)
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Petrovskis, I (författare)
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Dislers, A (författare)
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Ose, V (författare)
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Skrastina, D (författare)
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Gelderblom, HR (författare)
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- Lundkvist, A (författare)
- Karolinska Institutet
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Meisel, H (författare)
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Borisova, G (författare)
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Kruger, DH (författare)
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Pumpens, P (författare)
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Ulrich, R (författare)
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(creator_code:org_t)
- 2003-01-30
- 2002
- Engelska.
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Ingår i: Intervirology. - : S. Karger AG. - 0300-5526 .- 1423-0100. ; 45:4-6, s. 340-349
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http://kipublication...
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https://doi.org/10.1...
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Abstract
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- In recent years, epitopes of various origin have been inserted into the core protein of hepatitis B virus (HBc), allowing the formation of chimeric HBc particles. Although the C-terminus of a C-terminally truncated HBc (HBcΔ) tolerates the insertion of extended foreign sequences, the insertion capacity is still a limiting factor for the construction of multivalent vaccines. Previously, we described a new system to generate HBcΔ mosaic particles based on a read-through mechanism in an <i>Escherichia coli</i> suppressor strain [J Gen Virol 1997;78:2049–2053]. Those mosaic particles allowed the insertion of a 114-amino acid (aa)-long segment of a Puumala hantavirus (PUUV) nucleocapsid (N) protein. To study the value and the potential limitations of the mosaic approach in more detail, we investigated the assembly capacity of ‘non-mosaic’ HBcΔ fusion proteins and the corresponding mosaic constructs carrying 94, 213 and 433 aa of the hantaviral N protein. Whereas the fusion proteins carrying 94, 114, 213 or 433 aa were not assembled into HBcΔ particles, or only at a low yield, the insertion of a stop codon-bearing linker restored the ability to form particles with 94, 114 and 213 foreign aa. The mosaic particles formed exhibited PUUV-N protein antigenicity. Immunization of BALB/c mice with these mosaic particles carrying PUUV-N protein aa 1–114, aa 1–213 and aa 340–433, respectively, induced HBc-specific antibodies, whereas PUUV-N protein-specific antibodies were detected only in mice immunized with particles carrying N-terminal aa 1–114 or aa 1–213 of the N protein. Both the anti-HBc and anti-PUUV antibody responses were IgG1 dominated. In conclusion, stop codon suppression allows the formation of mosaic core particles carrying large-sized and ‘problematic’, e.g. hydrophobic, hantavirus sequences.
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- art (ämneskategori)
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Kazaks, A
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Lachmann, S
-
Koletzki, D
-
Petrovskis, I
-
Dislers, A
-
Ose, V
-
visa fler...
-
Skrastina, D
-
Gelderblom, HR
-
Lundkvist, A
-
Meisel, H
-
Borisova, G
-
Kruger, DH
-
Pumpens, P
-
Ulrich, R
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visa färre...
- Artiklar i publikationen
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Intervirology
- Av lärosätet
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Karolinska Institutet