Construction and Characterization of a Recombinant Invertebrate Iridovirus

A. Ozgen, H. Muratoglu, Z. Demirbag, J.M. Vlak, M.M. van Oers, R. Nalcacioglu

Research output: Contribution to journalArticleAcademicpeer-review

7 Citations (Scopus)

Abstract

Chilo iridescent virus (CIV), officially named Insect iridescent virus 6 (IIV6), is the type species of the genus Iridovirus (family Iridoviridae). In this paper we constructed a recombinant CIV, encoding the green fluorescent protein (GFP). This recombinant can be used to investigate viral replication dynamics. We showed that homologous recombination is a valid method to make CIV gene knockouts and to insert foreign genes. The CIV 157L gene, putatively encoding a non-functional inhibitor of apoptosis (IAP), was chosen as target for foreign gene insertion. The gfp open reading frame preceded by the viral mcp promoter was inserted into the 157L locus by homologous recombination in Anthonomus grandis BRL-AG-3A cells. Recombinant virus (rCIV-¿157L-gfp) was purified by successive rounds of plaque purification. All plaques produced by the purified recombinant virus emitted green fluorescence due to the presence of GFP. One-step growth curves for recombinant and wild-type CIV were similar and the recombinant was fully infectious in vivo. Hence, CIV157L can be inactivated without altering the replication kinetics of the virus. Consequently, the CIV 157L locus can be used as a site for insertion of foreign DNA, e.g. to modify viral properties for insect biocontrol.
Original languageEnglish
Pages (from-to)286-292
JournalVirus Research
Volume189
DOIs
Publication statusPublished - 2014

Keywords

  • chilo-iridescent-virus
  • complete genome sequence
  • membrane-protein chimera
  • major capsid protein
  • vaccinia virus
  • proteomic analysis
  • dna polymerase
  • blackfly larva
  • cell-surface
  • host-range

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