Purification and properties of the herpes simplex virus type 1 UL8 protein.
A rapid and simple two-step scheme for the purification of herpes simplex virus type 1 UL8 protein from insect cells infected with a recombinant baculovirus has been developed. The scheme involves DEAE-Sepharose and phenyl-Sepharose chromatography and yields approximately 1.5 mg of protein from 2.4 x 10(8) infected cells. The protein remains intact during purification as judged by its reactivity with amino and carboxy termini-specific antisera. Gel filtration chromatography showed that the protein exists as a monomer in solution. No binding of the protein to ssDNA or dsDNA or to a DNA/RNA hybrid could be demonstrated using a gel mobility shift assay.
Medical Research Council Virology Unit, Institute of Virology, Glasgow, U.K.
M E
ME
Parry
N D
ND
Stow
H S
HS
Marsden
D000595:Amino Acid Sequence; D000818:Animals; D016367:Baculoviridae; D001483:Base Sequence; D015153:Blotting, Western; D004265:DNA Helicases / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D019915:DNA Primase; D007313:Insecta; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000737:chemistry / Q000378:metabolism; D012316:RNA Nucleotidyltransferases / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D011994:Recombinant Proteins / Q000302:isolation & purification; D018139:Simplexvirus / Q000737:chemistry*; D014764:Viral Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism
D046911:Macromolecular Substances; D009838:Oligodeoxyribonucleotides; D011994:Recombinant Proteins; D014764:Viral Proteins; D019915:DNA Primase; D012316:RNA Nucleotidyltransferases; C062021:helicase-primase, Human herpesvirus 1; D004265:DNA Helicases