Characterization of the herpes simplex virus type 1 strain 17+ neurovirulence gene RL1 and its expression in a bacterial system.
The DNA sequence of herpes simplex virus type 1 (HSV-1) strain 17+ in the region coding for the polypeptide ICP34.5 predicts a protein of 248 amino acids with a proposed M(r) of 26,158. The entire RL1 open reading frame was cloned into the expression vector pET8c to enable over-expression of ICP34.5 in Escherichia coli. The expressed protein was partially purified and used as an immunogen to produce a polyclonal antiserum in rabbits. Construction of an ICP34.5 null mutant (1771), demonstrated that the predicted open reading frame for ICP34.5 in strain 17+ is correct and confirmed that HSV-1 strain 17+ ICP34.5 specifically determines neurovirulence. The specificity of the anti-serum directed against the E. coli-expressed ICP34.5 was defined by Western blotting of wild-type and RL1-negative infected cell extracts.
McKie
E A
EA
MRC Virology Unit, Institute of Virology, Glasgow, U.K.
Hope
R G
RG
Brown
S M
SM
MacLean
A R
AR
D000818:Animals; D000906:Antibodies; D001921:Brain / Q000382:microbiology*; D004279:DNA, Viral / Q000032:analysis; D005455:Fluorescent Antibody Technique; D005814:Genes, Viral / Q000235:genetics*; D018259:Herpesvirus 1, Human / Q000235:genetics* / Q000254:growth & development / Q000472:pathogenicity; D007928:Lethal Dose 50; D051379:Mice; D008807:Mice, Inbred BALB C; D008970:Molecular Weight; D009154:Mutation / Q000235:genetics; D014764:Viral Proteins / Q000032:analysis / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D014774:Virulence / Q000235:genetics
D000906:Antibodies; D004279:DNA, Viral; D014764:Viral Proteins; C066469:gamma 34.5 protein, Human herpesvirus 1