Archives of Virology

, Volume 141, Issue 12, pp 2457–2464 | Cite as

Functional expression of the bovine herpesvirus 1 alkaline deoxyribonuclease (UL12) inEscherichia coli

  • Y.-T. Chung
  • W. Hsu
Brief Report

Summary

Sequence analysis within the unique long segment of the bovine herpesvirus 1 (BHV-1; infectious bovine rhinotracheitis virus) genome identified an open reading frame whose deduced protein product of 487 amino acids exhibited homology to alkaline deoxyribonucleases (DNases) of other herpesviruses. To determine this BHV-1 gene product has nuclease activity, the gene designated UL12 was inserted into the vector pET-28a(+) and expressed inEscherichia coli as an oligohistidine-tagged protein. Upon induction with isopropyl β-D-thiogalactopyranosideE. coli BL21 (DE3) [pLysS] cells carrying this recombinant plasmid produced a 57-kDa protein, the molecular mass of which was in accordance with the prediction from the DNA sequence. The recombinant UL12 protein purified by nickel-chelating affinity chromatography exhibited both exonuclease and endonuclease activity, each with an alkaline pH optimum.

Keywords

Isopropyl Functional Expression Nuclease Activity Endonuclease Activity Deoxyribonuclease 

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Copyright information

© Springer-Verlag 1996

Authors and Affiliations

  • Y.-T. Chung
    • 1
  • W. Hsu
    • 1
  1. 1.Department of Veterinary MedicineNational Chung-Hsing UniversityTaichungTaiwan

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