Isolation and characterization of deletion mutants of herpes simplex virus type 2 (strain HG52).
We provide evidence that: (i) two variants lacking the XbaI site at map coordinate 0.7 have been selected following XbaI treatment of the DNA of herpes simplex virus type 2 strain HG52; (ii) one of these mutants had lost the 0.7 restriction site due to a deletion of approximately 150 base pairs and in the other the site loss was due to a similar sized sequence insertion; (iii) following XbaI treatment, four variants with deletions ranging in size from 1.5 kb (in both TRL and IRL) to 9 kb in IRL were isolated; (iv) substantial deletions in the long terminal repeat regions of HG52 are present with a frequency of 24% of genomes in the elite stock, a variant with a 3.75 kb deletion in IRL making up 10% and one with a 1.5 kb deletion in both IRL and TRL making up 14%; (v) two of the variants isolated after XbaI treatment of viral DNA were identical to the deletion prototype within the elite stock, suggesting that these variants were not generated as a result of XbaI treatment but pre-existed in the viral DNA pool; (vi) the deletion variants were stably maintained during routine stock propagation, were viable and could be propagated as cloned populations; (vii) the deletions did not have a marked deleterious effect on the one-step growth kinetics of the virus.
Harland
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Brown
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D001345:Autoradiography; D002872:Chromosome Deletion; D004262:DNA Restriction Enzymes; D004279:DNA, Viral / Q000032:analysis; D015245:Deoxyribonuclease BamHI; D015246:Deoxyribonuclease EcoRI; D015252:Deoxyribonucleases, Type II Site-Specific; D005333:Fetus; D005814:Genes, Viral; D006801:Humans; D008168:Lung; D009154:Mutation; D012091:Repetitive Sequences, Nucleic Acid; D018139:Simplexvirus / Q000235:genetics* / Q000254:growth & development / Q000302:isolation & purification; D014764:Viral Proteins / Q000096:biosynthesis; D014776:Virus Cultivation
D004279:DNA, Viral; D014764:Viral Proteins; D004262:DNA Restriction Enzymes; D015245:Deoxyribonuclease BamHI; D015246:Deoxyribonuclease EcoRI; C025580:endodeoxyribonuclease XBAI; D015252:Deoxyribonucleases, Type II Site-Specific