The molecular biology of human enteric caliciviruses
The molecular biology of human enteric caliciviruses
An authentic full length clone of Manchester SLV was constructed and used to undertake the first in vitro transcription and translation studies with a full length SLV. Transcription and translation studies of Manchester SLV revealed a proteolytic pattern which suggested the presence of a number of precursor proteins. Precursor proteins and mature proteins of Southampton NLV and Rabbit haemorrhagic disease virus had been mapped using polyclonal antisera to defined regions of the polyprotein. Defined regions of Manchester SLV were expressed in an E.coli expression system and the recombinant proteins purified and used to raise hyperimrnune antisera. These antisera were used in radio-immune precipitation studies, allowing identification of the processing products. In addition, the structural protein of Manchester SLV was expressed in insect cells using a baculovirus expression system. The structural proteins of Norwalk virus, and other characterised NLVs, when expressed in this system, produce recombinant protein which self assembles to produce virus-like particles (VLPs). VLPs were not observed after expression of Manchester SLV although a protein of the expected size was produced. This protein was used to generate polyclonal antisera, allowing detection of the structural protein from the in vitro processing products using a radio-immune precipitation assay. Six stool samples containing putative 'Sapporo-like' caliciviruses were obtained from a year long survey of non-bacterial gastroenteritis outbreaks in the South-West of England. Sequence obtained from four of these samples suggested two groups of 'Sapporo-like' caliciviruses present in the community. The second group of SLVs has only recently been recognised and oniy 3kb of the approximate 7.5kb genome had previously been sequenced. The complete genome of a group II SLV was sequenced during this study, which will allow further characterisation of these little known viruses.
University of Southampton
1999
Robinson, Samantha Jayne
(1999)
The molecular biology of human enteric caliciviruses.
University of Southampton, Doctoral Thesis.
Record type:
Thesis
(Doctoral)
Abstract
An authentic full length clone of Manchester SLV was constructed and used to undertake the first in vitro transcription and translation studies with a full length SLV. Transcription and translation studies of Manchester SLV revealed a proteolytic pattern which suggested the presence of a number of precursor proteins. Precursor proteins and mature proteins of Southampton NLV and Rabbit haemorrhagic disease virus had been mapped using polyclonal antisera to defined regions of the polyprotein. Defined regions of Manchester SLV were expressed in an E.coli expression system and the recombinant proteins purified and used to raise hyperimrnune antisera. These antisera were used in radio-immune precipitation studies, allowing identification of the processing products. In addition, the structural protein of Manchester SLV was expressed in insect cells using a baculovirus expression system. The structural proteins of Norwalk virus, and other characterised NLVs, when expressed in this system, produce recombinant protein which self assembles to produce virus-like particles (VLPs). VLPs were not observed after expression of Manchester SLV although a protein of the expected size was produced. This protein was used to generate polyclonal antisera, allowing detection of the structural protein from the in vitro processing products using a radio-immune precipitation assay. Six stool samples containing putative 'Sapporo-like' caliciviruses were obtained from a year long survey of non-bacterial gastroenteritis outbreaks in the South-West of England. Sequence obtained from four of these samples suggested two groups of 'Sapporo-like' caliciviruses present in the community. The second group of SLVs has only recently been recognised and oniy 3kb of the approximate 7.5kb genome had previously been sequenced. The complete genome of a group II SLV was sequenced during this study, which will allow further characterisation of these little known viruses.
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Published date: 1999
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Local EPrints ID: 464009
URI: http://eprints.soton.ac.uk/id/eprint/464009
PURE UUID: a0831bb5-d80d-407c-b7fa-3b75e2651762
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Date deposited: 04 Jul 2022 21:00
Last modified: 04 Jul 2022 21:00
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Author:
Samantha Jayne Robinson
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