title: Generation of an Avian-Mammalian Rotavirus Reassortant by Using a Helper Virus-Dependent Reverse Genetics System.
abstract: The genetic diversity of rotavirus A (RVA) strains is facilitated in part by genetic reassortment. Although this process of genome segment exchange has been reported frequently among mammalian RVAs, it remained unknown if mammalian RVAs also could package genome segments from avian RVA strains. We generated a simian RVA strain SA11 reassortant containing the VP4 gene of chicken RVA strain 02V0002G3. To achieve this, we transfected BSR5/T7 cells with a T7 polymerase-driven VP4-encoding plasmid, infected the cells with a temperature-sensitive SA11 VP4 mutant, and selected the recombinant virus by increasing the temperature. The reassortant virus could be stably passaged and exhibited cytopathic effects in MA-104 cells, but it replicated less efficiently than both parental viruses. Our results show that avian and mammalian rotaviruses can exchange genome segments, resulting in replication-competent reassortants with new genomic and antigenic features.
  
  
  OBJECTIVE
  This study shows that rotaviruses of mammals can package genome segments from rotaviruses of birds. The genetic diversity of rotaviruses could be broadened by this process, which might be important for their antigenic variability. The reverse genetics system applied in the study could be useful for targeted generation and subsequent characterization of distinct rotavirus reassortant strains.
authors.affiliation: Federal Institute for Risk Assessment, Berlin, Germany Reimar.Johne@bfr.bund.de.
authors.forename: Reimar
authors.initials: R
authors.lastname: Johne
authors.affiliation: Federal Institute for Risk Assessment, Berlin, Germany.
authors.forename: Jochen
authors.initials: J
authors.lastname: Reetz
authors.affiliation: Institute for Virology, Freie Universitaet Berlin, Berlin, Germany.
authors.forename: Benedikt B
authors.initials: BB
authors.lastname: Kaufer
authors.affiliation: Federal Institute for Risk Assessment, Berlin, Germany.
authors.forename: Eva
authors.initials: E
authors.lastname: Trojnar
mesh_terms: D000818:Animals; D036022:Capsid Proteins / Q000235:genetics; D002460:Cell Line; D002645:Chickens; D000882:Haplorhini; D006378:Helper Viruses / Q000235:genetics*; D016865:Reassortant Viruses / Q000235:genetics* / Q000254:growth & development*; D011995:Recombination, Genetic; D059386:Reverse Genetics / Q000379:methods*; D012401:Rotavirus / Q000235:genetics* / Q000254:growth & development* / Q000302:isolation & purification; D014776:Virus Cultivation
chemical_list: D036022:Capsid Proteins; C060339:VP4 protein, Rotavirus