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Sequencing and Characterisation of p74 Gene in Two Isolates of Anticarsia Gemmatalis MNPV

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Abstract

P74 is a protein encoded in the genome of baculoviruses, associated with the envelopes of occluded virus. Its presence proved to be essential for per os infection. In first place, in this work we designed two universal primers to amplify a sequence region of the p74 ORF in baculoviruses from different classification groups. Then, by the use of these amplicons we obtained the complete sequence of the p74 locus from two isolates of AgMNPV, 2D (Brazil) and SF (Argentina). In the flanking regions we determined the complete sequence of p10 gene and a portion of p26 gene. Comparing both p74 sequence data (ORFs of 1935 bp) we found fifteen nucleotide changes that result in six amino acid changes. Comparisons of AgMNPV p74s with other baculovirus homologous genes indicate a close relationship with other group I Nucleopolyhedrovirus, in particular CfDEFNPV. These results were based on ORF sequence, amino acid sequence and gene order. The predictive studies about secondary structure and hydrophobic index point at six regions potentially associated to its function or native conformation. Finally, the detection of p74 mRNA after virus DNA replication confirms a late expression pattern.

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Acknowledgements

This work was supported by grants from Agencia Nacional de Promoción Científica y Tecnológica (ANPCyT, FONCyT) and Universidad Nacional de Quilmes (UNQ) to PDG. MNB, MFB and MGP hold fellowships from CONICET, and VAR holds a fellowship from CIC-PBA. PDG and VR hold a research career award from CONICET (Consejo Nacional de Investigaciones Científicas y Técnicas, Argentina). ASC is a staff researcher at IMYZA-INTA (Instituto de Microbiología y Zoología Agrícola, Instituto Nacional de Tecnología Agropecuaria). We thank to BM Ribeiro for the AgMNPV-2D viral clone, JD Claus and S Lorenzatti for the AgMNPV-SF isolate, P Caballero for LdMNPV DNA and H Lauzon and B Arif for CfMNPV DNA.

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Authors and Affiliations

  1. LIGBCM (Laboratorio de Ingeniería Genética y Biología Celular y Molecular), Departamento Ciencia y Tecnología, Universidad Nacional de Quilmes, Roque Saenz Peña 180, Bernal (1876), Argentina

    Mariano Nicolás Belaich, Vanina Andrea Rodríguez, Marcos Fabián Bilen, Marcela Gabriela Pilloff, Victor Romanowski & Pablo Daniel Ghiringhelli

  2. IMYZA (Instituto de Microbiología y Zoología Agrícola), Instituto Nacional de Tecnología Agropecuaria, CC 25, Castelar (1712), Argentina

    Mariano Nicolás Belaich, Vanina Andrea Rodríguez & Alicia Sciocco-Cap

  3. IBBM (Instituto de Bioquímica y Biología Molecular), Facultad de Ciencias exactas y Naturales, Universidad Nacional de La Plata, 49 y 115, La Plata (1900), Argentina

    Victor Romanowski

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  1. Mariano Nicolás Belaich
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  2. Vanina Andrea Rodríguez
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  3. Marcos Fabián Bilen
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  7. Pablo Daniel Ghiringhelli
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Correspondence to Pablo Daniel Ghiringhelli.

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Belaich, M.N., Rodríguez, V.A., Bilen, M.F. et al. Sequencing and Characterisation of p74 Gene in Two Isolates of Anticarsia Gemmatalis MNPV. Virus Genes 32, 59–70 (2006). https://doi.org/10.1007/s11262-005-5846-z

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  • DOI: https://doi.org/10.1007/s11262-005-5846-z

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