Mapping and sequence of the gene for the pseudorabies virus glycoprotein which accumulates in the medium of infected cells

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Abstract
RNA from pseudorabies virus (PRV)-infected Vero cells was translated in a reticulocyte lysate with and without the addition of dog pancreas microsomes. Upon addition of the microsomes to the translation reaction, an additional prominent protein product was observed that was not present when microsomes were omitted. The gene coding for this processed protein and its lower MW precursor was mapped witin the small unique region of the genome by hybridization of mRNA to cloned fragments of PRV DNA and translation of the selected mRNA. A fragment of the coding region of this gene was inserted into an open reading frame cloning vector to express part of this gene as a hybrid protein in Escherichia coli. This hybrid protein was injected into mice to raise an antiserum which was found to precipitate the glycoprotein which accumulates in the medium of PRV-infected cells. Evidently, the gene for the excreted glycoprotein (gX) maps to the small unique region of the genome, and the precursor of this glycoprotein is readily processed by dog pancreas microsomes. The region of the PRV genome which codes for this glycoprotein was sequenced and found to include an open reading frame coding for 498 amino acids, flanked by sequences which contain features common to eukaryotic promoters and polyadenylation signals. The predicted protein sequence includes a hydrophobic sequence at the N-terminus which could be a signal sequence, and a hydrophobic sequence followed by a hydrophilic sequence at the C-terminus.