Premium
The UL2 Open Reading Frame of Bovine Herpesvirus 1 Encodes a Uracil‐DNA Glycosylase
Author(s) -
Chung YangTsung,
Hsu Weili
Publication year - 1996
Publication title -
microbiology and immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.664
H-Index - 70
eISSN - 1348-0421
pISSN - 0385-5600
DOI - 10.1111/j.1348-0421.1996.tb01164.x
Subject(s) - open reading frame , biology , uracil dna glycosylase , dna glycosylase , microbiology and biotechnology , peptide sequence , nucleic acid sequence , dna , biochemistry , escherichia coli , gene , dna damage
Sequence analysis within the unique long segment of the bovine herpesvirus 1 (BHV‐1) genome previously identified an open reading frame (ORF), designated UL2, whose deduced polypeptide of 204 amino acids contained a consensus uracil‐DNA glycosylase (UDGase) signature sequence. To determine whether the BHV‐1 UL2 ORF product has UDGase activity, we positioned the UL2 sequence downstream of the T7 promoter on the vector pET‐28b(+) and expressed it in Escherichia coli . Upon induction with isopropyl β‐ D ‐thiogalactopyranoside these cells produced a 23‐kDa protein, the molecular mass of which was in accordance with the prediction from the nucleotide sequence. A one‐step purification procedure using nickel‐chelating affinity chromatography resulted in a homogeneous preparation of this protein, which displayed specific UDGase activity in an in vitro enzyme assay. These results provide evidence that the BHV‐1 UL2 gene does encode a UDGase.