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Octamer and heat shock elements regulate transcription from the AcMNPV polyhedrin gene promoter

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Abstract

The baculovirus expression vector system exploits the polyhedrin (polh) promoter for high expression of foreign proteins in insect cells. The mechanism of basal and hyperactivated transcription from this promoter, however, remains poorly understood. We have analyzed the 4-kb upstream region of the polh promoter; deletion of two separate parts of the 4-kb upstream region, harboring the Oct binding site and the heat shock element, respectively, resulted in significant reduction of reporter gene expression regulated by the polh promoter. Insect cell host factors could bind to these elements in vitro. Moreover, these elements could activate polh transcription during viral infection when present upstream of a minimal polh promoter in transient expression reporter assays. Our results suggest the possible existence of transcription factors belonging to the POU and heat shock transcription factor family in Spodoptera frugiperda cells and support the hypothesis that host proteins may play a major role in activating transcription from the polh promoter.

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Acknowledgments

This work was partly supported by Department of Science and Technology Grant # HR/OY/GB-16/99 (to MDB) and by core support from the Department of Biotechnology, Government of India, to Centre for DNA Fingerprinting and Diagnostics. The authors wish to thank the National Genomics and Transcriptomics Facility at CDFD, Hyderabad, India, for DNA sequencing. AR and SK were recipients of Junior and Senior Research Fellowships from the Council for Scientific and Industrial Research, Government of India.

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Author notes

  1. M. Senthil Kumar

    Present address: Department of Molecular Microbiology and Infectious Diseases, College of Medicine, Florida International University, Miami, FL, 33199, USA

  2. Aruna Ramachandran

    Present address: Urology Department, Children’s Hospital Boston, Boston, MA, 02115, USA

Authors and Affiliations

  1. Laboratory of Molecular Oncology, Centre for DNA Fingerprinting and Diagnostics, Block no. 7, 5-4-399/B, Gruhakalpa complex, Nampally, Hyderabad, 500001, India

    M. Senthil Kumar & Murali Dharan Bashyam

  2. Laboratory of Molecular and Cellular Biology, Centre for DNA Fingerprinting and Diagnostics, Block no. 7, 5-4-399/B, Gruhakalpa complex, Nampally, Hyderabad, 500001, India

    M. Senthil Kumar, Aruna Ramachandran & Seyed E. Hasnain

  3. University of Hyderabad, Hyderabad, India

    Seyed E. Hasnain

  4. Jawaharlal Nehru Centre for Advanced Scientific Research, Bangalore, 560064, India

    Seyed E. Hasnain

Authors
  1. M. Senthil Kumar
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  2. Aruna Ramachandran
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  3. Seyed E. Hasnain
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  4. Murali Dharan Bashyam
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Correspondence to Murali Dharan Bashyam.

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Kumar, M.S., Ramachandran, A., Hasnain, S.E. et al. Octamer and heat shock elements regulate transcription from the AcMNPV polyhedrin gene promoter. Arch Virol 154, 445–456 (2009). https://doi.org/10.1007/s00705-009-0324-x

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  • DOI: https://doi.org/10.1007/s00705-009-0324-x

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