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High-level heterologous gene expression in Ochrobactrum anthropi using an A-rich UP element

  • Applied Genetics and Molecular Biotechnology
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Abstract

DNA regions that flank a gene’s promoter play an important role in determining transcription efficiency by interacting with the carboxy-terminal domain of RNA polymerase α-subunit. We placed an adenine-rich upstream element (UP) between −38 and −59 of the core trc promoter to enhance gene expression in Ochrobactrum anthropi up to 66-fold. The high level of expression achieved by the UP element and the N-terminus fusion of a 6xHis epitope tag facilitated detection and purification of heterologous proteins directly from O. anthropi.

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Acknowledgements

We express our appreciation to Dr. Tim Larson of Virginia Tech for his critical reading of the manuscript and suggestions. We would like to thank Mrs. Kay Carlson of Virginia tech for providing us with O. anthropi strain and the support throughout the experiment.

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Authors and Affiliations

  1. Department of Biomedical Sciences and Pathobiology, Center for Molecular Medicine and Infectious Diseases, Virginia-Maryland Regional College of Veterinary Medicine, Virginia Polytechnic Institute and State University, 1410 Prices Fork Rd, Blacksburg, VA, 24061-0342, USA

    Mohamed Seleem, Stephen M. Boyle & Nammalwar Sriranganathan

  2. Department of Forensic Medicine and Toxicology, Faculty of Veterinary Medicine, Assiut University, Assiut, Egypt

    Mohammed Ali

  3. Department of Microbiology, Faculty of Veterinary Medicine, South Valley University, Qena, Egypt

    M. W. Abd Al-Azeem

Authors
  1. Mohamed Seleem
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  2. Mohammed Ali
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  3. M. W. Abd Al-Azeem
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  4. Stephen M. Boyle
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  5. Nammalwar Sriranganathan
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Corresponding author

Correspondence to Nammalwar Sriranganathan.

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Seleem, M., Ali, M., Abd Al-Azeem, M.W. et al. High-level heterologous gene expression in Ochrobactrum anthropi using an A-rich UP element. Appl Microbiol Biotechnol 73, 1123–1127 (2007). https://doi.org/10.1007/s00253-006-0555-7

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  • DOI: https://doi.org/10.1007/s00253-006-0555-7

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