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Transcriptional analysis of ORF amv133 of Amsacta moorei entomopoxvirus

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Abstract

The open reading frame (ORF) amv133 of Amsacta moorei entomopoxvirus, encodes a putative lipase gene. Its temporal expression pattern was characterized by RT-PCR and found to start at 6 h postinfection (h p.i.) and reach a maximum level at 48 h p.i. While the ORF has a late promoter motif, the inhibition of viral DNA synthesis by Ara-C failed to inhibit transcription, but a general inhibitor of protein synthesis prevented its transcription, indicating that amv133 is an intermediate gene. 5′-RACE analysis showed that transcription was initiated at position −77 relative to the translational start site. To determine the size of the promoter, several truncations were generated and cloned upstream of the firefly luciferase reporter gene. The resulting constructs were tested in a dual assay. A fragment that contained 115 bp relative to the transcription start site exhibited optimum promoter length.

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Acknowledgments

The Scientific and Technological Research Council of Turkey, TÜBITAK, Project No. 109T975, supported this study financially. The authors would like to acknowledge Prof. Basil M Arif for his critical evaluation of the manuscript.

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  1. Department of Biology, Faculty of Science, Karadeniz Technical University, Trabzon, Turkey

    Emine Ozsahin, Kazım Sezen & Zihni Demirbag

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  1. Emine Ozsahin
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  2. Kazım Sezen
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  3. Zihni Demirbag
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Correspondence to Zihni Demirbag.

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Ozsahin, E., Sezen, K. & Demirbag, Z. Transcriptional analysis of ORF amv133 of Amsacta moorei entomopoxvirus. Arch Virol 159, 2541–2547 (2014). https://doi.org/10.1007/s00705-014-2096-1

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