Journal: Journal of Virology
Article Title: A Virus with a Mutation in the ICP4-Binding Site in the L/ST Promoter of Herpes Simplex Virus Type 1, but Not a Virus with a Mutation in Open Reading Frame P, Exhibits Cell-Type-Specific Expression of ?134.5 Transcripts and Latency-Associated Transcripts
Figure Lengend Snippet: Physical map of the internal repeat region of HSV-1 DNA. (A) Diagram of the HSV-1 genome. TR L , U L , and IR L , terminal repeat, unique, and internal repeat sequences, respectively, of the long segment; IR S , U S , and TR S , internal repeat, unique, and terminal repeat sequences, respectively, of the short segment; b′ , internal inverted repeat sequence bracketing U L ; c ′, internal inverted repeat sequence bracketing U S ; a′ , inverted sequence located between the b′ and c ′ sequences and, in reverse orientation, at the genomic termini. (B) Expanded map of the internal repeat sequence b′-a′-c′ lying between kbp 117 and 134 on the HSV-1 genome. Beneath the scale of kilobase pairs are shown the locations of the b′-a′-c′ sequence and oriS. (C) Beneath the b-a-c repeats are shown the locations of genes contained within kbp 117 to 134. Specifically, the map shows the locations of sequences specifying the transcripts encoding ICP0, ICP34.5, ICP4, and ICP22 and sequences specifying the LATs. ORFs, including ORF-O and ORF-P, are shown as hatched boxes beneath ORF34.5. The locations of sequences specifying the L/STs are shown beneath ORF34.5 and ORF-P. Beneath the L/STs are shown sequences specifying TR/UL RNA, the αX and βX transcripts, and ori S RNAs 1 and 2. Arrows indicate the direction of transcription, and the polyadenylation status is designated A + , A − or ? (D) DNA sequences specifying the riboprobes (arrows) used in this study. The arrows represent the orientation of these sequences in pGEM vectors as driven by the SP6 promoter. (E) Sequences specifying the DNA probes used for Southern blot analysis. (F to L) Physical map locations of the HSV-1 DNA sequences located in plasmids pBamK (F); pNco, pL/ST- n 38, and pL/ST-4BS m (G); pL/ST-Nxi and pL/ST-NxiLM3 (H); pWR-CAT + , pL/ST-U-CAT, and pL/ST-4BS m -CAT (heterologous CAT sequences are shown as boxes) (I); p0V, p0V- n 38, p0V-4BS, and p0V-4/38 (J); pG-L/ST and pG-L/ST-C (heterologous GST sequences are shown as boxes) (K); and p38Nco and p4BSNco (L). (M) Locations of the L/ST mutations constructed in this study. The position of the 4BS mutation is represented by the triangle. The position of the n 38 mutation is represented by the rectangle. Below the physical map is the wild-type (KOS) sequence, beneath which are the mutated (L/ST-4BS and L/ST- n 38) sequences. Each point mutation is designated by a vertical line. The Eco 47III restriction site generated by the mutation in L/ST-4BS and the Pfl MI restriction site destroyed by the L/ST- n 38 mutation are underlined.
Article Snippet: Riboprobes were transcribed from the SP6 promoter in the vector as instructed by the manufacturer (Promega).
Techniques: Sequencing, Southern Blot, Construct, Mutagenesis, Generated