Low Temperature-Dependent Salmonid Alphavirus Glycoprotein Processing and Recombinant Virus-Like Particle Formation
Pancreas disease (PD) and sleeping disease (SD) are important viral scourges in aquaculture of Atlantic salmon and rainbow trout. The etiological agent of PD and SD is salmonid alphavirus (SAV), an unusual member of the Togaviridae (genus Alphavirus). SAV replicates at lower temperatures in fish. Ou...
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ftciteseerx:oai:CiteSeerX.psu:10.1.1.289.4663 2023-05-15T15:32:27+02:00 Low Temperature-Dependent Salmonid Alphavirus Glycoprotein Processing and Recombinant Virus-Like Particle Formation Stefan W. Metz Femke Feenstra Stephane Villoing Marielle C. Van Hulten Jan W. Van Lent Joseph Koumans Just M. Vlak Gorben P. Pijlman The Pennsylvania State University CiteSeerX Archives 2011 application/zip http://citeseerx.ist.psu.edu/viewdoc/summary?doi=10.1.1.289.4663 en eng http://citeseerx.ist.psu.edu/viewdoc/summary?doi=10.1.1.289.4663 Metadata may be used without restrictions as long as the oai identifier remains attached to it. ftp://ftp.ncbi.nlm.nih.gov/pub/pmc/6c/f7/PLoS_One_2011_Oct_3_6(10)_e25816.tar.gz text 2011 ftciteseerx 2016-01-07T21:26:58Z Pancreas disease (PD) and sleeping disease (SD) are important viral scourges in aquaculture of Atlantic salmon and rainbow trout. The etiological agent of PD and SD is salmonid alphavirus (SAV), an unusual member of the Togaviridae (genus Alphavirus). SAV replicates at lower temperatures in fish. Outbreaks of SAV are associated with large economic losses of,17 to 50 million $/year. Current control strategies rely on vaccination with inactivated virus formulations that are cumbersome to obtain and have intrinsic safety risks. In this research we were able to obtain non-infectious virus-like particles (VLPs) of SAV via expression of recombinant baculoviruses encoding SAV capsid protein and two major immunodominant viral glycoproteins, E1 and E2 in Spodoptera frugiperda Sf9 insect cells. However, this was only achieved when a temperature shift from 27uC to lower temperatures was applied. At 27uC, precursor E2 (PE2) was misfolded and not processed by host furin into mature E2. Hence, E2 was detected neither on the surface of infected cells nor as VLPs in the culture fluid. However, when temperatures during protein expression were lowered, PE2 was processed into mature E2 in a temperaturedependent manner and VLPs were abundantly produced. So, temperature shift-down during synthesis is a prerequisite for Text Atlantic salmon Unknown Sav’ ENVELOPE(156.400,156.400,68.817,68.817) |
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English |
description |
Pancreas disease (PD) and sleeping disease (SD) are important viral scourges in aquaculture of Atlantic salmon and rainbow trout. The etiological agent of PD and SD is salmonid alphavirus (SAV), an unusual member of the Togaviridae (genus Alphavirus). SAV replicates at lower temperatures in fish. Outbreaks of SAV are associated with large economic losses of,17 to 50 million $/year. Current control strategies rely on vaccination with inactivated virus formulations that are cumbersome to obtain and have intrinsic safety risks. In this research we were able to obtain non-infectious virus-like particles (VLPs) of SAV via expression of recombinant baculoviruses encoding SAV capsid protein and two major immunodominant viral glycoproteins, E1 and E2 in Spodoptera frugiperda Sf9 insect cells. However, this was only achieved when a temperature shift from 27uC to lower temperatures was applied. At 27uC, precursor E2 (PE2) was misfolded and not processed by host furin into mature E2. Hence, E2 was detected neither on the surface of infected cells nor as VLPs in the culture fluid. However, when temperatures during protein expression were lowered, PE2 was processed into mature E2 in a temperaturedependent manner and VLPs were abundantly produced. So, temperature shift-down during synthesis is a prerequisite for |
author2 |
The Pennsylvania State University CiteSeerX Archives |
format |
Text |
author |
Stefan W. Metz Femke Feenstra Stephane Villoing Marielle C. Van Hulten Jan W. Van Lent Joseph Koumans Just M. Vlak Gorben P. Pijlman |
spellingShingle |
Stefan W. Metz Femke Feenstra Stephane Villoing Marielle C. Van Hulten Jan W. Van Lent Joseph Koumans Just M. Vlak Gorben P. Pijlman Low Temperature-Dependent Salmonid Alphavirus Glycoprotein Processing and Recombinant Virus-Like Particle Formation |
author_facet |
Stefan W. Metz Femke Feenstra Stephane Villoing Marielle C. Van Hulten Jan W. Van Lent Joseph Koumans Just M. Vlak Gorben P. Pijlman |
author_sort |
Stefan W. Metz |
title |
Low Temperature-Dependent Salmonid Alphavirus Glycoprotein Processing and Recombinant Virus-Like Particle Formation |
title_short |
Low Temperature-Dependent Salmonid Alphavirus Glycoprotein Processing and Recombinant Virus-Like Particle Formation |
title_full |
Low Temperature-Dependent Salmonid Alphavirus Glycoprotein Processing and Recombinant Virus-Like Particle Formation |
title_fullStr |
Low Temperature-Dependent Salmonid Alphavirus Glycoprotein Processing and Recombinant Virus-Like Particle Formation |
title_full_unstemmed |
Low Temperature-Dependent Salmonid Alphavirus Glycoprotein Processing and Recombinant Virus-Like Particle Formation |
title_sort |
low temperature-dependent salmonid alphavirus glycoprotein processing and recombinant virus-like particle formation |
publishDate |
2011 |
url |
http://citeseerx.ist.psu.edu/viewdoc/summary?doi=10.1.1.289.4663 |
long_lat |
ENVELOPE(156.400,156.400,68.817,68.817) |
geographic |
Sav’ |
geographic_facet |
Sav’ |
genre |
Atlantic salmon |
genre_facet |
Atlantic salmon |
op_source |
ftp://ftp.ncbi.nlm.nih.gov/pub/pmc/6c/f7/PLoS_One_2011_Oct_3_6(10)_e25816.tar.gz |
op_relation |
http://citeseerx.ist.psu.edu/viewdoc/summary?doi=10.1.1.289.4663 |
op_rights |
Metadata may be used without restrictions as long as the oai identifier remains attached to it. |
_version_ |
1766362952175190016 |