Rescue of a herpes simplex virus type 1 neurovirulence function with a cloned DNA fragment.
A herpes simplex virus type 1 (HSV-1) genetic function that is required for viral replication in the murine central nervous system was unambiguously localized. Thus, cosmid clones of either HSV-1 HindIII fragment C (0.64 to 0.87 map units) or fragment B (0.64 to 0.83 plus 0.91 to 1.0 map units) were employed to restore neurovirulence to an intertypic recombinant (RE6) that is specifically deficient in this property. The neurovirulent recombinants were generated in cell culture by cotransfecting the clone fragments and unit-length RE6 DNA and then selected in mouse brains. Either fragment efficiently conferred neurovirulence to RE6, demonstrating that no short region unique sequences are required. Analyses of the genomic structures of the neurovirulent recombinants showed that, in every case, HSV-1 information from 0.71 to 0.83 map units was incorporated into the RE6 genome. Cleavage of HindIII fragment C with EcoRI eliminated its capacity to rescue RE6. Virulence could be restored by the addition of HSV-1 BamHI fragment L (0.71 to 0.74 map units) that spans an EcoRI site at 0.72 map units. The precise location of this HSV-1 neurovirulence function is discussed.
Thompson
R L
RL
Devi-Rao
G V
GV
Stevens
J G
JG
Wagner
E K
EK
D000818:Animals; D001921:Brain / Q000382:microbiology*; D003001:Cloning, Molecular; D004262:DNA Restriction Enzymes; D004279:DNA, Viral; D015246:Deoxyribonuclease EcoRI; D015247:Deoxyribonuclease HindIII; D005814:Genes, Viral; D007928:Lethal Dose 50; D051379:Mice; D011995:Recombination, Genetic; D018139:Simplexvirus / Q000235:genetics / Q000472:pathogenicity*; D014162:Transfection; D014774:Virulence
D004279:DNA, Viral; D004262:DNA Restriction Enzymes; D015246:Deoxyribonuclease EcoRI; D015247:Deoxyribonuclease HindIII