High level expression and purification of herpes simplex virus type 1 immediate early polypeptide Vmw110.
Herpes simplex virus type 1 (HSV-1) encodes five immediate early (IE) polypeptides. This paper reports the construction of a baculovirus vector which expresses large amounts of Vmw110, the product of IE gene 1. The expressed protein has been purified to near homogeneity and has a mobility on SDS polyacrylamide gels identical to that of Vmw110 produced during HSV-1 infection. Characterisation of its properties indicated that it forms dimers and perhaps higher order oligomers in solution and that the purified protein binds to both single stranded and double stranded calf thymus DNA cellulose columns. However, filter binding experiments were unable to detect any stable association of Vmw110 with DNA in solution.
Everett
R D
RD
MRC Virology Unit, Glasgow, UK.
Orr
A
A
Elliott
M
M
D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D015153:Blotting, Western; D002417:Cattle; D002853:Chromatography, Liquid; D004247:DNA / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D015870:Gene Expression; D005814:Genes, Viral; D005822:Genetic Vectors; D017874:Immediate-Early Proteins; D018139:Simplexvirus / Q000378:metabolism*; D044767:Ubiquitin-Protein Ligases; D014764:Viral Proteins / Q000235:genetics* / Q000302:isolation & purification
D004268:DNA-Binding Proteins; C050678:IE1 protein, Human herpesvirus 1; D017874:Immediate-Early Proteins; D014764:Viral Proteins; D004247:DNA; D044767:Ubiquitin-Protein Ligases; C051804:Vmw110 protein, Human herpesvirus 1