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Functional and structural characterisation of AgMNPV ie1

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Abstract

We have located and cloned the Anticarsia gemmatalis multicapsid nucleopolyhedrovirus isolate 2D (AgMNPV-2D) genomic DNA fragment containing the immediate early 1 ORF and its flanking regions. Computer assisted analysis of the complete ie1 locus nucleotide sequence information was used to locate regulatory signals in the upstream region and conserved nucleotide and amino acid sequences. Comparative studies led to the identification of several characteristic protein motifs and to the conclusion that AgMNPV-2D is more closely related to Choristoneura fumiferana defective NPV than to other Group I nucleopolyhedrovirus. We have also shown that the AgMNPV IE1 protein was able to transactivate an early Autographa californica MNPV promoter and its own promoter in transient expression assays. In order to investigate the biological functionality of the ie1 promoter, the ie1 upstream activating region (UAR) was molecularly dissected and cloned upstream of the E. coli lacZ ORF. The results obtained, after transfection of UFL-AG-286 insect cells, leading us to find that the −492 and −357 versions contains sequence motifs important for the level of the lacZ reporter gene expression.

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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; Centro Argentino Brasileño de Biotecnología (CABBIO) to VR, PDG and BMR. MFB and MGP hold a fellowship from CONICET. PDG, VR and MEL are Research Career Members of CONICET, Argentina.

Author information

Correspondence to Pablo Daniel Ghiringhelli.

Additional information

The GenBank accession number of the sequence reported in this paper is AF368905.

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Bilen, M.F., Pilloff, M.G., Belaich, M.N. et al. Functional and structural characterisation of AgMNPV ie1 . Virus Genes 35, 549–562 (2007). https://doi.org/10.1007/s11262-007-0150-8

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Keywords

  • Baculovirus
  • Anticarsia gemmatalis
  • AgMNPV
  • ie1
  • Immediate early genes