Formation of virus-like particles from cloned cDNAs of Thogoto virus.
Thogoto virus (THOV) is the type species of tick-transmitted orthomyxoviruses. Here, we describe the generation of virus-like particles (VLP) of THOV from cloned cDNAs. To synthesize the six structural proteins of THOV in mammalian cells, we used T7-controlled expression plasmids and a recombinant vaccinia virus producing T7 RNA polymerase. A minireplicon encoding a reporter gene flanked by THOV promoter sequences was expressed by the cellular RNA polymerase I. The recombinant proteins were functional in encapsidation, amplification and transcription of the minireplicon RNA. Furthermore, the artificial nucleocapsids were packaged into THO-VLPs that transferred the minireplicon to indicator cells. This system should be helpful in generating recombinant THOV entirely from cloned cDNAs.
Wagner
Elke
E
Abteilung Virologie, Institut für Medizinische Mikrobiologie und Hygiene, Universität Freiburg, D-79008 Freiburg, Germany1.
Engelhardt
Othmar G
OG
Abteilung Virologie, Institut für Medizinische Mikrobiologie und Hygiene, Universität Freiburg, D-79008 Freiburg, Germany1.
Weber
Friedemann
F
Abteilung Virologie, Institut für Medizinische Mikrobiologie und Hygiene, Universität Freiburg, D-79008 Freiburg, Germany1.
Haller
Otto
O
Abteilung Virologie, Institut für Medizinische Mikrobiologie und Hygiene, Universität Freiburg, D-79008 Freiburg, Germany1.
Kochs
Georg
G
Abteilung Virologie, Institut für Medizinische Mikrobiologie und Hygiene, Universität Freiburg, D-79008 Freiburg, Germany1.
D000818:Animals; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D012321:DNA-Directed RNA Polymerases / Q000235:genetics / Q000378:metabolism; D017930:Genes, Reporter / Q000235:genetics; D005814:Genes, Viral / Q000235:genetics*; D007106:Immune Sera / Q000276:immunology / Q000494:pharmacology; D009500:Neutralization Tests; D011401:Promoter Regions, Genetic / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000378:metabolism; D018117:Thogotovirus / Q000187:drug effects / Q000235:genetics* / Q000276:immunology / Q000502:physiology*; D014162:Transfection; D014616:Vaccinia virus / Q000235:genetics; D014709:Vero Cells; D014764:Viral Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D019065:Virus Assembly
D018076:DNA, Complementary; D007106:Immune Sera; D011994:Recombinant Proteins; D014764:Viral Proteins; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases