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Applied Microbiology and Biotechnology

, Volume 73, Issue 5, pp 1123–1127 | Cite as

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

  • Mohamed Seleem
  • Mohammed Ali
  • M. W. Abd Al-Azeem
  • Stephen M. Boyle
  • Nammalwar SriranganathanEmail author
Applied Genetics and Molecular Biotechnology

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.

Keywords

Methylparathion Miller Unit Brucella Abortus Upstream Element Express Fusion Protein 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

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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Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  • Mohamed Seleem
    • 1
  • Mohammed Ali
    • 2
  • M. W. Abd Al-Azeem
    • 3
  • Stephen M. Boyle
    • 1
  • Nammalwar Sriranganathan
    • 1
    Email author
  1. 1.Department of Biomedical Sciences and Pathobiology, Center for Molecular Medicine and Infectious Diseases, Virginia-Maryland Regional College of Veterinary MedicineVirginia Polytechnic Institute and State UniversityBlacksburgUSA
  2. 2.Department of Forensic Medicine and Toxicology, Faculty of Veterinary MedicineAssiut UniversityAssiutEgypt
  3. 3.Department of Microbiology, Faculty of Veterinary MedicineSouth Valley UniversityQenaEgypt

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