Variations in Herpes Simplex Virus Isolated From Human Ganglia and a Study of Clonal Variation in Hsv‐1*

113

125

SUMMARYThe nucleotide sequence of 4 kilobases of DNA from within the short region of the genome of herpes simplex virus type 1 has been determined. This portion of DNA contains the junctions of the terminal and internal inverted repeat sequence components with the unique sequence component and the 5' regions of the genes which encode the Vmw 12, Vmw 68 and Vmw 175 immediate-early polypeptides. The transcription and translation initiation sites of all three genes and the 5' and 3' boundaries of the Vmw 12 and Vmw 68 gene introns have been localized on the DNA sequence and shown to be flanked by sequences which resemble those found in similar positions in other eukaryotic genes. For the Vmw 12 and Vmw 68 genes the promoters, the 5'-non-coding regions of the mRNAs, and the introns lie within the terminal and internal inverted repeats respectively while the polypeptide-coding regions lie in the short unique component. The introns consist largely of tandemly reiterated copies of a 22-nucleotide sequence: the Vmw 12 gene intron contains seven of these and the Vmw 68 gene intron five. The Vmw 175 gene is located entirely within the short repeats, of which those regions sequenced here have the extremely high G + C content of 78 %, in marked contrast to the value of 66 % obtained for the adjacent region of the unique sequence component. Prediction of the complete amino acid sequence of the Vmw 12 polypeptide, accounting for a mol. wt. of 9830, and of the first 523 amino-terminal amino acids of the Vmw 175 polypeptide has been made from the DNA sequence. The polypeptide-coding region of the Vmw 175 gene has an average G + C content of 80% but nevertheless specifies a wide range of amino acid types because of the maximal assignment of G and C residues to codon third base positions.

Section: Discussionmentioning

confidence: 99%

DNA Sequence Anaylysis of an Immediate-Early Gene Region of the Herpes Simplex Virus Type 1 Genome (Map Coordinates 0.950 to 0.978)

Murchie

McGeoch

1982

113

125

“…This is due to size heterogeneities in the repetitive regions of the genome and is not a consequence of the deletions in the BamHI p fragments (Lonsdale et al, 1980 …”

Section: Analysis Of Recombinant D N a Profiles From Purified Virion mentioning

confidence: 99%

Thymidine Kinase Deletion Mutants of Herpes Simplex Virus Type 1

Sanders

Wilkie

Davison

1982

277 SUMMARYDeletions in the cloned thymidine kinase (TK) gene of herpes simplex virus type 1 (HSV-I), strain 17 syn ÷, were produced by two methods. Removal of a 506 base pair fragment from between the unique SstI and BglII restriction endonuclease sites of pTK1 (HSV-1 BamHI p cloned in pAT153) and subsequent transformation of Escherichia coli resulted in the isolation of 50 deleted plasmids. Sequential digestion of pTK1 with BgllI and nuclease BAL 31 followed by ligation and recleavage with BglII resulted in the isolation of 31 deleted plasmids. Three clones, pTK2, pTK3 and pTK4, obtained following BglII and SstI treatment of pTK 1 were recombined with wild-type (wt) HSV-1 (17) syn ÷ DNA in baby hamster kidney (BHK) cells to produce TKdeletion mutants HSV-1 (17)TK 1301, HSV-1 (17)TK 1302 and HSV-1 (17)TK 1303 respectively. 5-Bromo-2'-deoxyuridine, 5-bromo-2'-deoxycytidine and 9-(2-hydroxyethoxymethyl)guanine were used to reduce the background of TK + virus in heterogeneous recombinant stocks analysed for the presence of TK-recombinants. All recombinant clones isolated produced a small syncytial plaque morphology in BHK cells. The mutants HSV-1 (17) TK 1301 and HSV-I (17) TK 1302 were TK-, failed to produce polypeptides of molecular weights 43 000 and 19 000 found in wt-infected cells and demonstrated one-step growth curves different from wt virus and the TK-mutant HSV-I (17)dPyk -7. Superinfection studies with HSV-1 (17)TK 1301, HSV-1 (17)TK 1302, HSV-I (MDK) and HSV-1 (17) dPyk -7 indicated that all TK-mutants except dPyK -7 produce a tram-acting gene product which can switch on the transforming HSV-1 TK gene.

“…Isolates USA29 and USA30 came from a trigeminal ganglion and the brain from a herpes encephalitis patient respectively and were obtained by Dr D. Lonsdale from Dr J. R. Baringer. The origin of the American and Japanese ganglion isolates is given in Lonsdale et al (1979Lonsdale et al ( , 1980. The origin of the genital isolates and the Saskatchewan facial isolates was reported by Chaney et al (1983), who made earlier restriction enzyme site comparisons; their data covered 53 of the 87 restriction endonuclease sites for these isolates, but the full characterization is included for completeness.…”

Section: Hsv-1 Isolates and Re Analysismentioning

confidence: 99%

Variable Restriction Endonuclease Sites of Herpes Simplex Virus Type 1 Isolates from Encephalitic, Facial and Genital Lesions and Ganglia

Chaney¹,

Warren

Subak‐Sharpe³

1983

Self Cite

SUMMARYThe distribution of restriction endonuclease (RE) sites was compared for 84 herpes simplex virus type 1 (HSV-1) isolates obtained from the ganglia, facial lesions, genital lesions and from brain tissue from herpes encephalitis cases. The isolates came from Canada, the U.K., the U.S.A. and Japan. Out of a total of 224 sites identified, 87 were variable. Three of the 30 most variable sites were at significantly (P < 0-05) different frequencies in groups of isolates from distinct anatomical sites of isolation; one of these, and a further two sites, were at significantly different frequencies in groups from distinct geographical origins. There are at least two inter-related linkage groups. However, most of the site combinations appear to be random. The variability of RE sites in contiguous genome segments, which include both non-coding and coding sequences, show a marked heterogeneity, indicating that some viral gene sequences are more variable than others. The three RE sites at different frequencies in viral groups from distinct anatomical sites of isolation are in two genome segments : map units 27 to 35 and 50 to 57. We infer from the observed associations with anatomical site of viral isolation that part of at least one of these segments may modulate viral virulence in man following infection.