sp6 promoter Search Results


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  • 93
    Thermo Fisher sp 6 promoter primer
    Sp 6 Promoter Primer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 39 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sp 6 promoter primer/product/Thermo Fisher
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    92
    Promega sp6 promoter
    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 <t>SP6</t> 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.
    Sp6 Promoter, supplied by Promega, used in various techniques. Bioz Stars score: 92/100, based on 573 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Thermo Fisher sp6 promoter
    Electrophoretic mobility of in vitro circularized Ty1 RNA. ( A ) <t>SP6</t>
    Sp6 Promoter, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 795 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Roche sp6 promoter
    Electrophoretic mobility of in vitro circularized Ty1 RNA. ( A ) <t>SP6</t>
    Sp6 Promoter, supplied by Roche, used in various techniques. Bioz Stars score: 92/100, based on 188 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    GE Healthcare sp6 promoter
    Electrophoretic mobility of in vitro circularized Ty1 RNA. ( A ) <t>SP6</t>
    Sp6 Promoter, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 92/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Promega sp6 promoter primers
    Analysis of TFIID–TFIIA binding to the natural AdML promoter. ( A ) DNase I footprinting reactions were performed with probes containing the AdML promoter (lanes 1–3,7–9 ) and the AdML(+1G,+3G) mutant promoter (lanes 4–6,10–12 ). Reactions contained no protein (lanes 1,4,7,10 ), 0.2 (lanes 2,3,5,6 ) or 0.5 (lanes 9,10,11,12 ) μl concentrated TFIID, and recombinant TFIIA (0.1 μl, lanes 3,6,9,12 ). The locations of the TATA and Inr elements, as well as regions of downstream protection (DS) and 4 hypersensitive sites (arrows) are depicted to the right. ( B ) In vitro transcription reactions were performed in HeLa nuclear extracts with supercoiled plasmids containing the AdML core promoter from −35 to +40 (lane 1; see Materials and Methods), the AdML core promoter containing two substitution mutations (+1G and +3G) within the Inr element (lane 2 ), the synthetic TATA–Inr core promoter (lane 3 ) and the synthetic TATA–Inr(+3G) core promoter (lane 4 ). Plasmid DNA (300 ng) and 100 μg HeLa nuclear extract were used for each reaction. RNA products were analyzed by primer extension, by use of the same radiolabeled <t>SP6</t> primer to insure that the cDNA products possessed similar specific activities.
    Sp6 Promoter Primers, supplied by Promega, used in various techniques. Bioz Stars score: 90/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sp6 promoter primers/product/Promega
    Average 90 stars, based on 17 article reviews
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    85
    Promega sp6 promoter oligonucleotide
    Analysis of TFIID–TFIIA binding to the natural AdML promoter. ( A ) DNase I footprinting reactions were performed with probes containing the AdML promoter (lanes 1–3,7–9 ) and the AdML(+1G,+3G) mutant promoter (lanes 4–6,10–12 ). Reactions contained no protein (lanes 1,4,7,10 ), 0.2 (lanes 2,3,5,6 ) or 0.5 (lanes 9,10,11,12 ) μl concentrated TFIID, and recombinant TFIIA (0.1 μl, lanes 3,6,9,12 ). The locations of the TATA and Inr elements, as well as regions of downstream protection (DS) and 4 hypersensitive sites (arrows) are depicted to the right. ( B ) In vitro transcription reactions were performed in HeLa nuclear extracts with supercoiled plasmids containing the AdML core promoter from −35 to +40 (lane 1; see Materials and Methods), the AdML core promoter containing two substitution mutations (+1G and +3G) within the Inr element (lane 2 ), the synthetic TATA–Inr core promoter (lane 3 ) and the synthetic TATA–Inr(+3G) core promoter (lane 4 ). Plasmid DNA (300 ng) and 100 μg HeLa nuclear extract were used for each reaction. RNA products were analyzed by primer extension, by use of the same radiolabeled <t>SP6</t> primer to insure that the cDNA products possessed similar specific activities.
    Sp6 Promoter Oligonucleotide, supplied by Promega, used in various techniques. Bioz Stars score: 85/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sp6 promoter oligonucleotide/product/Promega
    Average 85 stars, based on 12 article reviews
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    88
    Thermo Fisher sp6 dual promoters
    Analysis of TFIID–TFIIA binding to the natural AdML promoter. ( A ) DNase I footprinting reactions were performed with probes containing the AdML promoter (lanes 1–3,7–9 ) and the AdML(+1G,+3G) mutant promoter (lanes 4–6,10–12 ). Reactions contained no protein (lanes 1,4,7,10 ), 0.2 (lanes 2,3,5,6 ) or 0.5 (lanes 9,10,11,12 ) μl concentrated TFIID, and recombinant TFIIA (0.1 μl, lanes 3,6,9,12 ). The locations of the TATA and Inr elements, as well as regions of downstream protection (DS) and 4 hypersensitive sites (arrows) are depicted to the right. ( B ) In vitro transcription reactions were performed in HeLa nuclear extracts with supercoiled plasmids containing the AdML core promoter from −35 to +40 (lane 1; see Materials and Methods), the AdML core promoter containing two substitution mutations (+1G and +3G) within the Inr element (lane 2 ), the synthetic TATA–Inr core promoter (lane 3 ) and the synthetic TATA–Inr(+3G) core promoter (lane 4 ). Plasmid DNA (300 ng) and 100 μg HeLa nuclear extract were used for each reaction. RNA products were analyzed by primer extension, by use of the same radiolabeled <t>SP6</t> primer to insure that the cDNA products possessed similar specific activities.
    Sp6 Dual Promoters, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 88/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher t7 sp6 promoter
    Analysis of TFIID–TFIIA binding to the natural AdML promoter. ( A ) DNase I footprinting reactions were performed with probes containing the AdML promoter (lanes 1–3,7–9 ) and the AdML(+1G,+3G) mutant promoter (lanes 4–6,10–12 ). Reactions contained no protein (lanes 1,4,7,10 ), 0.2 (lanes 2,3,5,6 ) or 0.5 (lanes 9,10,11,12 ) μl concentrated TFIID, and recombinant TFIIA (0.1 μl, lanes 3,6,9,12 ). The locations of the TATA and Inr elements, as well as regions of downstream protection (DS) and 4 hypersensitive sites (arrows) are depicted to the right. ( B ) In vitro transcription reactions were performed in HeLa nuclear extracts with supercoiled plasmids containing the AdML core promoter from −35 to +40 (lane 1; see Materials and Methods), the AdML core promoter containing two substitution mutations (+1G and +3G) within the Inr element (lane 2 ), the synthetic TATA–Inr core promoter (lane 3 ) and the synthetic TATA–Inr(+3G) core promoter (lane 4 ). Plasmid DNA (300 ng) and 100 μg HeLa nuclear extract were used for each reaction. RNA products were analyzed by primer extension, by use of the same radiolabeled <t>SP6</t> primer to insure that the cDNA products possessed similar specific activities.
    T7 Sp6 Promoter, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    TaKaRa sp6 promoter plasmid
    Analysis of TFIID–TFIIA binding to the natural AdML promoter. ( A ) DNase I footprinting reactions were performed with probes containing the AdML promoter (lanes 1–3,7–9 ) and the AdML(+1G,+3G) mutant promoter (lanes 4–6,10–12 ). Reactions contained no protein (lanes 1,4,7,10 ), 0.2 (lanes 2,3,5,6 ) or 0.5 (lanes 9,10,11,12 ) μl concentrated TFIID, and recombinant TFIIA (0.1 μl, lanes 3,6,9,12 ). The locations of the TATA and Inr elements, as well as regions of downstream protection (DS) and 4 hypersensitive sites (arrows) are depicted to the right. ( B ) In vitro transcription reactions were performed in HeLa nuclear extracts with supercoiled plasmids containing the AdML core promoter from −35 to +40 (lane 1; see Materials and Methods), the AdML core promoter containing two substitution mutations (+1G and +3G) within the Inr element (lane 2 ), the synthetic TATA–Inr core promoter (lane 3 ) and the synthetic TATA–Inr(+3G) core promoter (lane 4 ). Plasmid DNA (300 ng) and 100 μg HeLa nuclear extract were used for each reaction. RNA products were analyzed by primer extension, by use of the same radiolabeled <t>SP6</t> primer to insure that the cDNA products possessed similar specific activities.
    Sp6 Promoter Plasmid, supplied by TaKaRa, used in various techniques. Bioz Stars score: 85/100, based on 20 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Cellfree Sciences sp6 promoter sequence
    Analysis of TFIID–TFIIA binding to the natural AdML promoter. ( A ) DNase I footprinting reactions were performed with probes containing the AdML promoter (lanes 1–3,7–9 ) and the AdML(+1G,+3G) mutant promoter (lanes 4–6,10–12 ). Reactions contained no protein (lanes 1,4,7,10 ), 0.2 (lanes 2,3,5,6 ) or 0.5 (lanes 9,10,11,12 ) μl concentrated TFIID, and recombinant TFIIA (0.1 μl, lanes 3,6,9,12 ). The locations of the TATA and Inr elements, as well as regions of downstream protection (DS) and 4 hypersensitive sites (arrows) are depicted to the right. ( B ) In vitro transcription reactions were performed in HeLa nuclear extracts with supercoiled plasmids containing the AdML core promoter from −35 to +40 (lane 1; see Materials and Methods), the AdML core promoter containing two substitution mutations (+1G and +3G) within the Inr element (lane 2 ), the synthetic TATA–Inr core promoter (lane 3 ) and the synthetic TATA–Inr(+3G) core promoter (lane 4 ). Plasmid DNA (300 ng) and 100 μg HeLa nuclear extract were used for each reaction. RNA products were analyzed by primer extension, by use of the same radiolabeled <t>SP6</t> primer to insure that the cDNA products possessed similar specific activities.
    Sp6 Promoter Sequence, supplied by Cellfree Sciences, used in various techniques. Bioz Stars score: 90/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sp6 promoter sequence/product/Cellfree Sciences
    Average 90 stars, based on 13 article reviews
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    85
    Promega sp6 promoter primer pairs
    Analysis of TFIID–TFIIA binding to the natural AdML promoter. ( A ) DNase I footprinting reactions were performed with probes containing the AdML promoter (lanes 1–3,7–9 ) and the AdML(+1G,+3G) mutant promoter (lanes 4–6,10–12 ). Reactions contained no protein (lanes 1,4,7,10 ), 0.2 (lanes 2,3,5,6 ) or 0.5 (lanes 9,10,11,12 ) μl concentrated TFIID, and recombinant TFIIA (0.1 μl, lanes 3,6,9,12 ). The locations of the TATA and Inr elements, as well as regions of downstream protection (DS) and 4 hypersensitive sites (arrows) are depicted to the right. ( B ) In vitro transcription reactions were performed in HeLa nuclear extracts with supercoiled plasmids containing the AdML core promoter from −35 to +40 (lane 1; see Materials and Methods), the AdML core promoter containing two substitution mutations (+1G and +3G) within the Inr element (lane 2 ), the synthetic TATA–Inr core promoter (lane 3 ) and the synthetic TATA–Inr(+3G) core promoter (lane 4 ). Plasmid DNA (300 ng) and 100 μg HeLa nuclear extract were used for each reaction. RNA products were analyzed by primer extension, by use of the same radiolabeled <t>SP6</t> primer to insure that the cDNA products possessed similar specific activities.
    Sp6 Promoter Primer Pairs, supplied by Promega, used in various techniques. Bioz Stars score: 85/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sp6 promoter primer pairs/product/Promega
    Average 85 stars, based on 7 article reviews
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    sp6 promoter primer pairs - by Bioz Stars, 2020-08
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    93
    Eurofins sp6 promoter
    Analysis of TFIID–TFIIA binding to the natural AdML promoter. ( A ) DNase I footprinting reactions were performed with probes containing the AdML promoter (lanes 1–3,7–9 ) and the AdML(+1G,+3G) mutant promoter (lanes 4–6,10–12 ). Reactions contained no protein (lanes 1,4,7,10 ), 0.2 (lanes 2,3,5,6 ) or 0.5 (lanes 9,10,11,12 ) μl concentrated TFIID, and recombinant TFIIA (0.1 μl, lanes 3,6,9,12 ). The locations of the TATA and Inr elements, as well as regions of downstream protection (DS) and 4 hypersensitive sites (arrows) are depicted to the right. ( B ) In vitro transcription reactions were performed in HeLa nuclear extracts with supercoiled plasmids containing the AdML core promoter from −35 to +40 (lane 1; see Materials and Methods), the AdML core promoter containing two substitution mutations (+1G and +3G) within the Inr element (lane 2 ), the synthetic TATA–Inr core promoter (lane 3 ) and the synthetic TATA–Inr(+3G) core promoter (lane 4 ). Plasmid DNA (300 ng) and 100 μg HeLa nuclear extract were used for each reaction. RNA products were analyzed by primer extension, by use of the same radiolabeled <t>SP6</t> primer to insure that the cDNA products possessed similar specific activities.
    Sp6 Promoter, supplied by Eurofins, used in various techniques. Bioz Stars score: 93/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sp6 promoter/product/Eurofins
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    91
    MWG-Biotech sp6 t7 promoter sequences
    Analysis of TFIID–TFIIA binding to the natural AdML promoter. ( A ) DNase I footprinting reactions were performed with probes containing the AdML promoter (lanes 1–3,7–9 ) and the AdML(+1G,+3G) mutant promoter (lanes 4–6,10–12 ). Reactions contained no protein (lanes 1,4,7,10 ), 0.2 (lanes 2,3,5,6 ) or 0.5 (lanes 9,10,11,12 ) μl concentrated TFIID, and recombinant TFIIA (0.1 μl, lanes 3,6,9,12 ). The locations of the TATA and Inr elements, as well as regions of downstream protection (DS) and 4 hypersensitive sites (arrows) are depicted to the right. ( B ) In vitro transcription reactions were performed in HeLa nuclear extracts with supercoiled plasmids containing the AdML core promoter from −35 to +40 (lane 1; see Materials and Methods), the AdML core promoter containing two substitution mutations (+1G and +3G) within the Inr element (lane 2 ), the synthetic TATA–Inr core promoter (lane 3 ) and the synthetic TATA–Inr(+3G) core promoter (lane 4 ). Plasmid DNA (300 ng) and 100 μg HeLa nuclear extract were used for each reaction. RNA products were analyzed by primer extension, by use of the same radiolabeled <t>SP6</t> primer to insure that the cDNA products possessed similar specific activities.
    Sp6 T7 Promoter Sequences, supplied by MWG-Biotech, used in various techniques. Bioz Stars score: 91/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Cellfree Sciences sp6 promoter
    Analysis of TFIID–TFIIA binding to the natural AdML promoter. ( A ) DNase I footprinting reactions were performed with probes containing the AdML promoter (lanes 1–3,7–9 ) and the AdML(+1G,+3G) mutant promoter (lanes 4–6,10–12 ). Reactions contained no protein (lanes 1,4,7,10 ), 0.2 (lanes 2,3,5,6 ) or 0.5 (lanes 9,10,11,12 ) μl concentrated TFIID, and recombinant TFIIA (0.1 μl, lanes 3,6,9,12 ). The locations of the TATA and Inr elements, as well as regions of downstream protection (DS) and 4 hypersensitive sites (arrows) are depicted to the right. ( B ) In vitro transcription reactions were performed in HeLa nuclear extracts with supercoiled plasmids containing the AdML core promoter from −35 to +40 (lane 1; see Materials and Methods), the AdML core promoter containing two substitution mutations (+1G and +3G) within the Inr element (lane 2 ), the synthetic TATA–Inr core promoter (lane 3 ) and the synthetic TATA–Inr(+3G) core promoter (lane 4 ). Plasmid DNA (300 ng) and 100 μg HeLa nuclear extract were used for each reaction. RNA products were analyzed by primer extension, by use of the same radiolabeled <t>SP6</t> primer to insure that the cDNA products possessed similar specific activities.
    Sp6 Promoter, supplied by Cellfree Sciences, used in various techniques. Bioz Stars score: 92/100, based on 28 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sp6 promoter/product/Cellfree Sciences
    Average 92 stars, based on 28 article reviews
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    Image Search Results


    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.

    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

    doi:

    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

    Electrophoretic mobility of in vitro circularized Ty1 RNA. ( A ) SP6

    Journal: RNA

    Article Title: An evaluation of detection methods for large lariat RNAs

    doi: 10.1261/rna.7124405

    Figure Lengend Snippet: Electrophoretic mobility of in vitro circularized Ty1 RNA. ( A ) SP6

    Article Snippet: Full-length Ty1 sense control RNA was synthesized from the SP6 promoter of linearized plasmid pCEC24 (Ty1 nucleotides 241–5463) using the Ambion MAXI-script in vitro transcription kit.

    Techniques: In Vitro

    Analysis of TFIID–TFIIA binding to the natural AdML promoter. ( A ) DNase I footprinting reactions were performed with probes containing the AdML promoter (lanes 1–3,7–9 ) and the AdML(+1G,+3G) mutant promoter (lanes 4–6,10–12 ). Reactions contained no protein (lanes 1,4,7,10 ), 0.2 (lanes 2,3,5,6 ) or 0.5 (lanes 9,10,11,12 ) μl concentrated TFIID, and recombinant TFIIA (0.1 μl, lanes 3,6,9,12 ). The locations of the TATA and Inr elements, as well as regions of downstream protection (DS) and 4 hypersensitive sites (arrows) are depicted to the right. ( B ) In vitro transcription reactions were performed in HeLa nuclear extracts with supercoiled plasmids containing the AdML core promoter from −35 to +40 (lane 1; see Materials and Methods), the AdML core promoter containing two substitution mutations (+1G and +3G) within the Inr element (lane 2 ), the synthetic TATA–Inr core promoter (lane 3 ) and the synthetic TATA–Inr(+3G) core promoter (lane 4 ). Plasmid DNA (300 ng) and 100 μg HeLa nuclear extract were used for each reaction. RNA products were analyzed by primer extension, by use of the same radiolabeled SP6 primer to insure that the cDNA products possessed similar specific activities.

    Journal: Genes & Development

    Article Title: Mechanism of synergy between TATA and initiator: synergistic binding of TFIID following a putative TFIIA-induced isomerization

    doi:

    Figure Lengend Snippet: Analysis of TFIID–TFIIA binding to the natural AdML promoter. ( A ) DNase I footprinting reactions were performed with probes containing the AdML promoter (lanes 1–3,7–9 ) and the AdML(+1G,+3G) mutant promoter (lanes 4–6,10–12 ). Reactions contained no protein (lanes 1,4,7,10 ), 0.2 (lanes 2,3,5,6 ) or 0.5 (lanes 9,10,11,12 ) μl concentrated TFIID, and recombinant TFIIA (0.1 μl, lanes 3,6,9,12 ). The locations of the TATA and Inr elements, as well as regions of downstream protection (DS) and 4 hypersensitive sites (arrows) are depicted to the right. ( B ) In vitro transcription reactions were performed in HeLa nuclear extracts with supercoiled plasmids containing the AdML core promoter from −35 to +40 (lane 1; see Materials and Methods), the AdML core promoter containing two substitution mutations (+1G and +3G) within the Inr element (lane 2 ), the synthetic TATA–Inr core promoter (lane 3 ) and the synthetic TATA–Inr(+3G) core promoter (lane 4 ). Plasmid DNA (300 ng) and 100 μg HeLa nuclear extract were used for each reaction. RNA products were analyzed by primer extension, by use of the same radiolabeled SP6 primer to insure that the cDNA products possessed similar specific activities.

    Article Snippet: For all probes, the radiolabeled downstream primer was the SP6 promoter primer (Promega, Inc.).

    Techniques: Binding Assay, Footprinting, Mutagenesis, Recombinant, In Vitro, Plasmid Preparation