Studies on caliciviral replication
Studies on caliciviral replication
This work reports the findings from studies on the replication of Jena virus (JV), a bovine norovirus discovered in 1984 in Jena, Germany. This particular virus displays similar tissue tropism and symptoms to the human-associated noroviruses and is thus a suitable candidate for an animal model of human disease. Analysis of JV protein synthesis in vitro revealed that a protein representative of the JV N-terminal polyprotein, detected in other noroviruses, is not apparently synthesised from JV. The hypothesis that this 5’ genomic region functions as an enhancer of translation was investigated by means of a novel EGFP/lacZ bi-cistronic expression system. The 5’ specific immunological reagents determines that this region is translated into protein, both in vitro and within a cell culture system, although the exact characteristics of this protein remain to be elucidated. These results also suggest that the proteolytic processing of the JV non-structural polyprotein is more complex than proteolytic events currently documented for other noroviruses, and indicate that the development of further immunological reagents is required for further analysis and identification of proteolytic products.
An attempt to propagate JV from a cDNA clone in an organ culture system using fresh bovine intestinal tissue is described. Preliminary data suggested that the viral capsid protein was synthesised but further optimisation of this system is required to demonstrate conclusive and repeatable results.
Currently, it is not known what factors prevent the replication of enteric noroviruses in cell culture or if a specific stage in the replicative cycle is blocked in standard laboratory tissue culture cells. To address this question an attempt was made to detect negative-sense JV RNA using an antisense reporter system. Negative-sense RNA was not detected using this technique, suggesting that standard cell lines do not support transcription of the infecting viral genome into negative stranded viral RNA.
University of Southampton
Salim, Omar
3b0bd7cd-bef2-4d17-86a7-9318054bc846
2005
Salim, Omar
3b0bd7cd-bef2-4d17-86a7-9318054bc846
Salim, Omar
(2005)
Studies on caliciviral replication.
University of Southampton, Doctoral Thesis.
Record type:
Thesis
(Doctoral)
Abstract
This work reports the findings from studies on the replication of Jena virus (JV), a bovine norovirus discovered in 1984 in Jena, Germany. This particular virus displays similar tissue tropism and symptoms to the human-associated noroviruses and is thus a suitable candidate for an animal model of human disease. Analysis of JV protein synthesis in vitro revealed that a protein representative of the JV N-terminal polyprotein, detected in other noroviruses, is not apparently synthesised from JV. The hypothesis that this 5’ genomic region functions as an enhancer of translation was investigated by means of a novel EGFP/lacZ bi-cistronic expression system. The 5’ specific immunological reagents determines that this region is translated into protein, both in vitro and within a cell culture system, although the exact characteristics of this protein remain to be elucidated. These results also suggest that the proteolytic processing of the JV non-structural polyprotein is more complex than proteolytic events currently documented for other noroviruses, and indicate that the development of further immunological reagents is required for further analysis and identification of proteolytic products.
An attempt to propagate JV from a cDNA clone in an organ culture system using fresh bovine intestinal tissue is described. Preliminary data suggested that the viral capsid protein was synthesised but further optimisation of this system is required to demonstrate conclusive and repeatable results.
Currently, it is not known what factors prevent the replication of enteric noroviruses in cell culture or if a specific stage in the replicative cycle is blocked in standard laboratory tissue culture cells. To address this question an attempt was made to detect negative-sense JV RNA using an antisense reporter system. Negative-sense RNA was not detected using this technique, suggesting that standard cell lines do not support transcription of the infecting viral genome into negative stranded viral RNA.
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Published date: 2005
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Local EPrints ID: 465616
URI: http://eprints.soton.ac.uk/id/eprint/465616
PURE UUID: 2bd7df64-b1fd-4308-bf8c-3ffabbcf0582
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Date deposited: 05 Jul 2022 02:04
Last modified: 23 Jul 2022 02:16
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Author:
Omar Salim
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