Detection of herpes simplex virus type 1 gene expression in latently and productively infected mouse ganglia using the polymerase chain reaction.
The polymerase chain reaction (PCR) was employed to detect herpes simplex virus (HSV) sequences in the DNA, and HSV gene expression in total cell RNA, extracted from cervical and trigeminal ganglia of mice during productive and latent infection with HSV-1, strain SC16. Such gene expression was detected in 1 microgram or less of RNA, the quantity anticipated to be present in one or two cervical ganglia. Within the limits of the primers available, gene expression during latency appeared to be restricted to the latency-associated transcript (LAT). The 195 base portion of the LAT amplified by the PCR was sequenced and found to contain several base changes and deletions with respect to published sequences for different HSV strains. These mutations, within the putative open reading frame 2 of the LAT, formed stop or terminator signals, which suggests that the LAT does not act to establish or maintain latency through translation to a protein. The primers for the LAT also amplified a 300 bp fragment from any murine and some other mammalian RNAs. Apart from the oligonucleotide primers, this fragment did not show any homology with HSV.
Lynas
C
C
Department of Ophthalmology, Medical School, Bristol, U.K.
Laycock
K A
KA
Cook
S D
SD
Hill
T J
TJ
Blyth
W A
WA
Maitland
N J
NJ
D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D004247:DNA / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics; D004259:DNA-Directed DNA Polymerase; D005260:Female; D005728:Ganglia, Sympathetic / Q000382:microbiology*; D005784:Gene Amplification; D005814:Genes, Viral; D007438:Introns; D051379:Mice; D008815:Mice, Inbred Strains; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D018139:Simplexvirus / Q000235:genetics*; D014158:Transcription, Genetic; D012668:Trigeminal Ganglion / Q000382:microbiology*; D014276:Trigeminal Nerve / Q000382:microbiology*
D004279:DNA, Viral; D015345:Oligonucleotide Probes; D004247:DNA; D004259:DNA-Directed DNA Polymerase