Expression of parvovirus LuIII NS1 from a Sindbis replicon for production of LuIII-luciferase transducing virus.
In order to develop an alternative packaging system for recombinant parvoviruses, the gene for the major nonstructural protein (NS1) of parvovirus LuIII was inserted into a Sindbis replicon vector. Cells infected with recombinant SinNS1 virus produced NS1 RNA from the Sindbis 26S promoter and expressed NS1 protein which was able to transactivate a parvovirus P38 promoter. Co-transfections of Sindbis-NS1 RNA together with a packageable LuIII transducing genome and a coat protein expression plasmid generated detectable levels of LuIII-luciferase transducing virus. These levels could be increased by a capsid expression plasmid that was also capable of expressing NS2. These results show that a multi-functional parvovirus protein expressed from a Sindbis RNA molecule can be used to produce recombinant parvoviruses.
Corsini
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Department of Microbiology, Colorado State University, Fort Collins 80523, USA. jcarlson@lamar.colostate.edu
Maxwell
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Maxwell
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Carlson
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D005822:Genetic Vectors / Q000378:metabolism; D008156:Luciferases / Q000235:genetics*; D017992:Parvovirus / Q000201:enzymology* / Q000235:genetics* / Q000502:physiology; D011401:Promoter Regions, Genetic; D012367:RNA, Viral / Q000096:biosynthesis; D012093:Replicon; D012845:Sindbis Virus / Q000235:genetics* / Q000378:metabolism; D015533:Transcriptional Activation; D014162:Transfection; D017361:Viral Nonstructural Proteins / Q000096:biosynthesis* / Q000235:genetics* / Q000502:physiology; D019065:Virus Assembly
D012367:RNA, Viral; D017361:Viral Nonstructural Proteins; D008156:Luciferases