title: Genome analysis and detection of a Chilean isolate of Grapevine leafroll associated virus-3.
abstract: The complete genome of the Chilean isolate Cl-766 of Grapevine leafroll-associated virus-3 (GLRaV-3) has been sequenced. This is the first genome sequence obtained from a GLRaV-3 isolate of the Southern hemisphere. The genomic RNA of 17,919 nucleotides contains 13 open reading frames (ORFs) with 5' and 3' untranslated regions (UTR) of 158 and 277 nucleotides, respectively. Comparison with NY1, the only isolate with complete genomic sequence available today, shows 97.6% nucleotide identity between the two isolates. Examination of the genome variability shows that most of the genetic diversity is concentrated in ORF1a. Three additional isolates from different geographic regions of Chile were partially sequenced as well, one which showed sequence divergence with respect to the other local and foreign isolates, indicative of different evolutionary constrains. Immunodetection systems were developed using monoclonal and polyclonal antibodies produced against the recombinant major coat protein of GLRaV-3, providing sensitive and specific detection using a triple antibody sandwich-enzyme linked immunosorbent assay (TAS-ELISA) and an immunocapture-reverse transcription-polymerase chain reaction (IC-RT-PCR) assay.
authors.affiliation: Fundación Ciencia para la Vida and Instituto Milenio de Biología Fundamental y Aplicada, Zañartu, 1482 Santiago, Chile, eaengeli@yahoo.com
authors.forename: Esteban A
authors.initials: EA
authors.lastname: Engel
authors.forename: Cristobal
authors.initials: C
authors.lastname: Girardi
authors.forename: Paula F
authors.initials: PF
authors.lastname: Escobar
authors.forename: Vania
authors.initials: V
authors.lastname: Arredondo
authors.forename: Calixto
authors.initials: C
authors.lastname: Domínguez
authors.forename: Tomás
authors.initials: T
authors.lastname: Pérez-Acle
authors.forename: Pablo D T
authors.initials: PD
authors.lastname: Valenzuela
mesh_terms: D000595:Amino Acid Sequence; D002677:Chile; D029381:Closteroviridae / Q000145:classification / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D016679:Genome, Viral; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010802:Phylogeny; D010935:Plant Diseases / Q000821:virology*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D014764:Viral Proteins / Q000032:analysis / Q000378:metabolism; D014773:Virology / Q000379:methods*; D027843:Vitis / Q000821:virology*
chemical_list: D014764:Viral Proteins