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  • 99
    Thermo Fisher expand high fidelity pcr system
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    Mechanisms of <t>GILZ</t> downregulation upon TLR activation. (A,B) AMs pretreated with BAY-11-7082 (BAY82, 5 μM), BAY-11-7085 (BAY85, 5 μM), or the solvent control DMSO (0.1%) for 1 h, followed by treatment with LPS (100 ng/mL), Pam 3 CSK 4 (Pam, 1 μg/mL), Poly(I:C) (PIC, 10 μg/mL), or medium (Co) for 4 h. GILZ expression was determined by Western blot. Tubulin served as a loading control. (A) Representative blot. (B) GILZ signal intensities were quantified and normalized to tubulin values ( n = 7). Values for unstimulated DMSO controls were set as 100%. (C,D) After preincubation with BAY-11-7082 or BAY-11-7085 (5 μM, 1 h), solvent (0.1% DMSO) or medium only (Co), AMs were treated with LPS (100 ng/mL) for 2 h. GILZ and ZFP36 mRNA expression was determined by <t>qRT-PCR</t> using ACTB as a housekeeping gene ( n = 3, duplicates). (E) AMs were either left untreated (Co) or treated with LPS (100 ng/mL), Pam 3 CSK 4 (Pam, 1 μg/mL), or Poly(I:C) (PIC, 10 μg/mL) for 4 h. TTP levels were determined by Western blot using tubulin as a loading control. GILZ signal intensities were normalized to tubulin and are shown as a percentage of untreated cells ( n = 2, triplicates). (F) The number of miRNAs predicted to target GILZ, CXCL8, IL6 , and TNF was assessed via the microRNA Data Integration Portal (mirDIP, accession date 02/02/2018). (G) AMs were either left untreated (Co) or treated with Poly(I:C) (PIC, 10 μg/mL), aurintricarboxylic acid (ATA, 25 μM), or a combination of both for 8 h. GILZ expression was quantified by Western blot. Signal intensities were normalized to tubulin and expressed as a percentage of untreated cells ( n = 3). * p
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    <t>PCR</t> amplification of <t>5-kbp</t> EAV-HP provirus products with putative pol region sequences. (A) Schematic diagram showing the positions of oligonucleotide primers EVJFOR and 103ER used for PCR relative to a complete provirus. The sequence recognized by 103ER is deleted from the 4-kbp provirus types, indicated as chicken EAV-HP (ev/J), but is present in the type IV ev/J clone 4-1 sequence. Ψ, packaging signal; SD, splice donor; SA, splice acceptor. (B) Ethidium bromide-stained agarose gel of separated PCR products amplified from line 21 chicken, RJF, and SJF DNA. The 0.6-kbp product from RJF was amplified from the type IV provirus, and a 5-kbp product with putative pol sequences was amplified only from SJF DNA.
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    <t>PCR</t> amplification of <t>5-kbp</t> EAV-HP provirus products with putative pol region sequences. (A) Schematic diagram showing the positions of oligonucleotide primers EVJFOR and 103ER used for PCR relative to a complete provirus. The sequence recognized by 103ER is deleted from the 4-kbp provirus types, indicated as chicken EAV-HP (ev/J), but is present in the type IV ev/J clone 4-1 sequence. Ψ, packaging signal; SD, splice donor; SA, splice acceptor. (B) Ethidium bromide-stained agarose gel of separated PCR products amplified from line 21 chicken, RJF, and SJF DNA. The 0.6-kbp product from RJF was amplified from the type IV provirus, and a 5-kbp product with putative pol sequences was amplified only from SJF DNA.
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    <t>PCR</t> amplification of <t>5-kbp</t> EAV-HP provirus products with putative pol region sequences. (A) Schematic diagram showing the positions of oligonucleotide primers EVJFOR and 103ER used for PCR relative to a complete provirus. The sequence recognized by 103ER is deleted from the 4-kbp provirus types, indicated as chicken EAV-HP (ev/J), but is present in the type IV ev/J clone 4-1 sequence. Ψ, packaging signal; SD, splice donor; SA, splice acceptor. (B) Ethidium bromide-stained agarose gel of separated PCR products amplified from line 21 chicken, RJF, and SJF DNA. The 0.6-kbp product from RJF was amplified from the type IV provirus, and a 5-kbp product with putative pol sequences was amplified only from SJF DNA.
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    <t>PCR</t> amplification of <t>5-kbp</t> EAV-HP provirus products with putative pol region sequences. (A) Schematic diagram showing the positions of oligonucleotide primers EVJFOR and 103ER used for PCR relative to a complete provirus. The sequence recognized by 103ER is deleted from the 4-kbp provirus types, indicated as chicken EAV-HP (ev/J), but is present in the type IV ev/J clone 4-1 sequence. Ψ, packaging signal; SD, splice donor; SA, splice acceptor. (B) Ethidium bromide-stained agarose gel of separated PCR products amplified from line 21 chicken, RJF, and SJF DNA. The 0.6-kbp product from RJF was amplified from the type IV provirus, and a 5-kbp product with putative pol sequences was amplified only from SJF DNA.
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    Genomic organization of tomato LeFPS genes. A, Thirty micrograms of tomato genomic <t>DNA</t> was digested with the indicated restriction endonuclease and subjected to DNA gel-blot analysis using <t>LeFPS1</t> cDNA as a hybridization probe. The blot was exposed for 4 d. Numbers on the left indicate size in kb from the markers (1-kb ladder, Gibco BRL, Cleveland). B, Partial map of the chromosomes 10 and 12. The putative location of the two loci (thick line) was deduced from the introgression line showing a polymorphism (dotted lines). LeFPS1 and LeFPS2 were mapped using the 1.3-kb LeFPS1 cDNA insert as a hybridization probe. The 0.18-kb cDNA fragment corresponding to the LeFPS1 3′-UTR was used to locate LeFPS1 on chromosome 12.
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    Genomic organization of tomato LeFPS genes. A, Thirty micrograms of tomato genomic <t>DNA</t> was digested with the indicated restriction endonuclease and subjected to DNA gel-blot analysis using <t>LeFPS1</t> cDNA as a hybridization probe. The blot was exposed for 4 d. Numbers on the left indicate size in kb from the markers (1-kb ladder, Gibco BRL, Cleveland). B, Partial map of the chromosomes 10 and 12. The putative location of the two loci (thick line) was deduced from the introgression line showing a polymorphism (dotted lines). LeFPS1 and LeFPS2 were mapped using the 1.3-kb LeFPS1 cDNA insert as a hybridization probe. The 0.18-kb cDNA fragment corresponding to the LeFPS1 3′-UTR was used to locate LeFPS1 on chromosome 12.
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    Genomic organization of tomato LeFPS genes. A, Thirty micrograms of tomato genomic <t>DNA</t> was digested with the indicated restriction endonuclease and subjected to DNA gel-blot analysis using <t>LeFPS1</t> cDNA as a hybridization probe. The blot was exposed for 4 d. Numbers on the left indicate size in kb from the markers (1-kb ladder, Gibco BRL, Cleveland). B, Partial map of the chromosomes 10 and 12. The putative location of the two loci (thick line) was deduced from the introgression line showing a polymorphism (dotted lines). LeFPS1 and LeFPS2 were mapped using the 1.3-kb LeFPS1 cDNA insert as a hybridization probe. The 0.18-kb cDNA fragment corresponding to the LeFPS1 3′-UTR was used to locate LeFPS1 on chromosome 12.
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    Genomic organization of tomato LeFPS genes. A, Thirty micrograms of tomato genomic <t>DNA</t> was digested with the indicated restriction endonuclease and subjected to DNA gel-blot analysis using <t>LeFPS1</t> cDNA as a hybridization probe. The blot was exposed for 4 d. Numbers on the left indicate size in kb from the markers (1-kb ladder, Gibco BRL, Cleveland). B, Partial map of the chromosomes 10 and 12. The putative location of the two loci (thick line) was deduced from the introgression line showing a polymorphism (dotted lines). LeFPS1 and LeFPS2 were mapped using the 1.3-kb LeFPS1 cDNA insert as a hybridization probe. The 0.18-kb cDNA fragment corresponding to the LeFPS1 3′-UTR was used to locate LeFPS1 on chromosome 12.
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    Genomic organization of tomato LeFPS genes. A, Thirty micrograms of tomato genomic <t>DNA</t> was digested with the indicated restriction endonuclease and subjected to DNA gel-blot analysis using <t>LeFPS1</t> cDNA as a hybridization probe. The blot was exposed for 4 d. Numbers on the left indicate size in kb from the markers (1-kb ladder, Gibco BRL, Cleveland). B, Partial map of the chromosomes 10 and 12. The putative location of the two loci (thick line) was deduced from the introgression line showing a polymorphism (dotted lines). LeFPS1 and LeFPS2 were mapped using the 1.3-kb LeFPS1 cDNA insert as a hybridization probe. The 0.18-kb cDNA fragment corresponding to the LeFPS1 3′-UTR was used to locate LeFPS1 on chromosome 12.
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    Genomic organization of tomato LeFPS genes. A, Thirty micrograms of tomato genomic <t>DNA</t> was digested with the indicated restriction endonuclease and subjected to DNA gel-blot analysis using <t>LeFPS1</t> cDNA as a hybridization probe. The blot was exposed for 4 d. Numbers on the left indicate size in kb from the markers (1-kb ladder, Gibco BRL, Cleveland). B, Partial map of the chromosomes 10 and 12. The putative location of the two loci (thick line) was deduced from the introgression line showing a polymorphism (dotted lines). LeFPS1 and LeFPS2 were mapped using the 1.3-kb LeFPS1 cDNA insert as a hybridization probe. The 0.18-kb cDNA fragment corresponding to the LeFPS1 3′-UTR was used to locate LeFPS1 on chromosome 12.
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    Genomic organization of tomato LeFPS genes. A, Thirty micrograms of tomato genomic <t>DNA</t> was digested with the indicated restriction endonuclease and subjected to DNA gel-blot analysis using <t>LeFPS1</t> cDNA as a hybridization probe. The blot was exposed for 4 d. Numbers on the left indicate size in kb from the markers (1-kb ladder, Gibco BRL, Cleveland). B, Partial map of the chromosomes 10 and 12. The putative location of the two loci (thick line) was deduced from the introgression line showing a polymorphism (dotted lines). LeFPS1 and LeFPS2 were mapped using the 1.3-kb LeFPS1 cDNA insert as a hybridization probe. The 0.18-kb cDNA fragment corresponding to the LeFPS1 3′-UTR was used to locate LeFPS1 on chromosome 12.
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    Genomic organization of tomato LeFPS genes. A, Thirty micrograms of tomato genomic <t>DNA</t> was digested with the indicated restriction endonuclease and subjected to DNA gel-blot analysis using <t>LeFPS1</t> cDNA as a hybridization probe. The blot was exposed for 4 d. Numbers on the left indicate size in kb from the markers (1-kb ladder, Gibco BRL, Cleveland). B, Partial map of the chromosomes 10 and 12. The putative location of the two loci (thick line) was deduced from the introgression line showing a polymorphism (dotted lines). LeFPS1 and LeFPS2 were mapped using the 1.3-kb LeFPS1 cDNA insert as a hybridization probe. The 0.18-kb cDNA fragment corresponding to the LeFPS1 3′-UTR was used to locate LeFPS1 on chromosome 12.
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    Genomic organization of tomato LeFPS genes. A, Thirty micrograms of tomato genomic <t>DNA</t> was digested with the indicated restriction endonuclease and subjected to DNA gel-blot analysis using <t>LeFPS1</t> cDNA as a hybridization probe. The blot was exposed for 4 d. Numbers on the left indicate size in kb from the markers (1-kb ladder, Gibco BRL, Cleveland). B, Partial map of the chromosomes 10 and 12. The putative location of the two loci (thick line) was deduced from the introgression line showing a polymorphism (dotted lines). LeFPS1 and LeFPS2 were mapped using the 1.3-kb LeFPS1 cDNA insert as a hybridization probe. The 0.18-kb cDNA fragment corresponding to the LeFPS1 3′-UTR was used to locate LeFPS1 on chromosome 12.
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    Genomic organization of tomato LeFPS genes. A, Thirty micrograms of tomato genomic <t>DNA</t> was digested with the indicated restriction endonuclease and subjected to DNA gel-blot analysis using <t>LeFPS1</t> cDNA as a hybridization probe. The blot was exposed for 4 d. Numbers on the left indicate size in kb from the markers (1-kb ladder, Gibco BRL, Cleveland). B, Partial map of the chromosomes 10 and 12. The putative location of the two loci (thick line) was deduced from the introgression line showing a polymorphism (dotted lines). LeFPS1 and LeFPS2 were mapped using the 1.3-kb LeFPS1 cDNA insert as a hybridization probe. The 0.18-kb cDNA fragment corresponding to the LeFPS1 3′-UTR was used to locate LeFPS1 on chromosome 12.
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    Genomic organization of tomato LeFPS genes. A, Thirty micrograms of tomato genomic <t>DNA</t> was digested with the indicated restriction endonuclease and subjected to DNA gel-blot analysis using <t>LeFPS1</t> cDNA as a hybridization probe. The blot was exposed for 4 d. Numbers on the left indicate size in kb from the markers (1-kb ladder, Gibco BRL, Cleveland). B, Partial map of the chromosomes 10 and 12. The putative location of the two loci (thick line) was deduced from the introgression line showing a polymorphism (dotted lines). LeFPS1 and LeFPS2 were mapped using the 1.3-kb LeFPS1 cDNA insert as a hybridization probe. The 0.18-kb cDNA fragment corresponding to the LeFPS1 3′-UTR was used to locate LeFPS1 on chromosome 12.
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    Genomic organization of tomato LeFPS genes. A, Thirty micrograms of tomato genomic <t>DNA</t> was digested with the indicated restriction endonuclease and subjected to DNA gel-blot analysis using <t>LeFPS1</t> cDNA as a hybridization probe. The blot was exposed for 4 d. Numbers on the left indicate size in kb from the markers (1-kb ladder, Gibco BRL, Cleveland). B, Partial map of the chromosomes 10 and 12. The putative location of the two loci (thick line) was deduced from the introgression line showing a polymorphism (dotted lines). LeFPS1 and LeFPS2 were mapped using the 1.3-kb LeFPS1 cDNA insert as a hybridization probe. The 0.18-kb cDNA fragment corresponding to the LeFPS1 3′-UTR was used to locate LeFPS1 on chromosome 12.
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    Genomic organization of tomato LeFPS genes. A, Thirty micrograms of tomato genomic <t>DNA</t> was digested with the indicated restriction endonuclease and subjected to DNA gel-blot analysis using <t>LeFPS1</t> cDNA as a hybridization probe. The blot was exposed for 4 d. Numbers on the left indicate size in kb from the markers (1-kb ladder, Gibco BRL, Cleveland). B, Partial map of the chromosomes 10 and 12. The putative location of the two loci (thick line) was deduced from the introgression line showing a polymorphism (dotted lines). LeFPS1 and LeFPS2 were mapped using the 1.3-kb LeFPS1 cDNA insert as a hybridization probe. The 0.18-kb cDNA fragment corresponding to the LeFPS1 3′-UTR was used to locate LeFPS1 on chromosome 12.
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    Image Search Results


    Mechanisms of GILZ downregulation upon TLR activation. (A,B) AMs pretreated with BAY-11-7082 (BAY82, 5 μM), BAY-11-7085 (BAY85, 5 μM), or the solvent control DMSO (0.1%) for 1 h, followed by treatment with LPS (100 ng/mL), Pam 3 CSK 4 (Pam, 1 μg/mL), Poly(I:C) (PIC, 10 μg/mL), or medium (Co) for 4 h. GILZ expression was determined by Western blot. Tubulin served as a loading control. (A) Representative blot. (B) GILZ signal intensities were quantified and normalized to tubulin values ( n = 7). Values for unstimulated DMSO controls were set as 100%. (C,D) After preincubation with BAY-11-7082 or BAY-11-7085 (5 μM, 1 h), solvent (0.1% DMSO) or medium only (Co), AMs were treated with LPS (100 ng/mL) for 2 h. GILZ and ZFP36 mRNA expression was determined by qRT-PCR using ACTB as a housekeeping gene ( n = 3, duplicates). (E) AMs were either left untreated (Co) or treated with LPS (100 ng/mL), Pam 3 CSK 4 (Pam, 1 μg/mL), or Poly(I:C) (PIC, 10 μg/mL) for 4 h. TTP levels were determined by Western blot using tubulin as a loading control. GILZ signal intensities were normalized to tubulin and are shown as a percentage of untreated cells ( n = 2, triplicates). (F) The number of miRNAs predicted to target GILZ, CXCL8, IL6 , and TNF was assessed via the microRNA Data Integration Portal (mirDIP, accession date 02/02/2018). (G) AMs were either left untreated (Co) or treated with Poly(I:C) (PIC, 10 μg/mL), aurintricarboxylic acid (ATA, 25 μM), or a combination of both for 8 h. GILZ expression was quantified by Western blot. Signal intensities were normalized to tubulin and expressed as a percentage of untreated cells ( n = 3). * p

    Journal: Frontiers in Immunology

    Article Title: Amplified Host Defense by Toll-Like Receptor-Mediated Downregulation of the Glucocorticoid-Induced Leucine Zipper (GILZ) in Macrophages

    doi: 10.3389/fimmu.2018.03111

    Figure Lengend Snippet: Mechanisms of GILZ downregulation upon TLR activation. (A,B) AMs pretreated with BAY-11-7082 (BAY82, 5 μM), BAY-11-7085 (BAY85, 5 μM), or the solvent control DMSO (0.1%) for 1 h, followed by treatment with LPS (100 ng/mL), Pam 3 CSK 4 (Pam, 1 μg/mL), Poly(I:C) (PIC, 10 μg/mL), or medium (Co) for 4 h. GILZ expression was determined by Western blot. Tubulin served as a loading control. (A) Representative blot. (B) GILZ signal intensities were quantified and normalized to tubulin values ( n = 7). Values for unstimulated DMSO controls were set as 100%. (C,D) After preincubation with BAY-11-7082 or BAY-11-7085 (5 μM, 1 h), solvent (0.1% DMSO) or medium only (Co), AMs were treated with LPS (100 ng/mL) for 2 h. GILZ and ZFP36 mRNA expression was determined by qRT-PCR using ACTB as a housekeeping gene ( n = 3, duplicates). (E) AMs were either left untreated (Co) or treated with LPS (100 ng/mL), Pam 3 CSK 4 (Pam, 1 μg/mL), or Poly(I:C) (PIC, 10 μg/mL) for 4 h. TTP levels were determined by Western blot using tubulin as a loading control. GILZ signal intensities were normalized to tubulin and are shown as a percentage of untreated cells ( n = 2, triplicates). (F) The number of miRNAs predicted to target GILZ, CXCL8, IL6 , and TNF was assessed via the microRNA Data Integration Portal (mirDIP, accession date 02/02/2018). (G) AMs were either left untreated (Co) or treated with Poly(I:C) (PIC, 10 μg/mL), aurintricarboxylic acid (ATA, 25 μM), or a combination of both for 8 h. GILZ expression was quantified by Western blot. Signal intensities were normalized to tubulin and expressed as a percentage of untreated cells ( n = 3). * p

    Article Snippet: Human GILZ 3′UTR cDNA was amplified using the Expand High fidelity PCR System (Sigma-Aldrich, # 11732641001) and the following primers: 5′-GCC TAC TAG TGC AGA GCC ACT AAA CTT G-3′ and 5′-AAT AGA GCT CAC TCT CAC AAA ACC CGC TAC-3′.

    Techniques: Activation Assay, Affinity Magnetic Separation, Expressing, Western Blot, Quantitative RT-PCR

    PCR amplification of 5-kbp EAV-HP provirus products with putative pol region sequences. (A) Schematic diagram showing the positions of oligonucleotide primers EVJFOR and 103ER used for PCR relative to a complete provirus. The sequence recognized by 103ER is deleted from the 4-kbp provirus types, indicated as chicken EAV-HP (ev/J), but is present in the type IV ev/J clone 4-1 sequence. Ψ, packaging signal; SD, splice donor; SA, splice acceptor. (B) Ethidium bromide-stained agarose gel of separated PCR products amplified from line 21 chicken, RJF, and SJF DNA. The 0.6-kbp product from RJF was amplified from the type IV provirus, and a 5-kbp product with putative pol sequences was amplified only from SJF DNA.

    Journal: Journal of Virology

    Article Title: Intact EAV-HP Endogenous Retrovirus in Sonnerat's Jungle Fowl

    doi: 10.1128/JVI.75.4.2029-2032.2001

    Figure Lengend Snippet: PCR amplification of 5-kbp EAV-HP provirus products with putative pol region sequences. (A) Schematic diagram showing the positions of oligonucleotide primers EVJFOR and 103ER used for PCR relative to a complete provirus. The sequence recognized by 103ER is deleted from the 4-kbp provirus types, indicated as chicken EAV-HP (ev/J), but is present in the type IV ev/J clone 4-1 sequence. Ψ, packaging signal; SD, splice donor; SA, splice acceptor. (B) Ethidium bromide-stained agarose gel of separated PCR products amplified from line 21 chicken, RJF, and SJF DNA. The 0.6-kbp product from RJF was amplified from the type IV provirus, and a 5-kbp product with putative pol sequences was amplified only from SJF DNA.

    Article Snippet: PCR was repeated using the Expand high-fidelity PCR system (Boehringer Mannheim) for cloning the 5-kbp SJF EAV-HP PCR product into the pGEM-T vector.

    Techniques: Polymerase Chain Reaction, Amplification, Sequencing, Staining, Agarose Gel Electrophoresis

    PCR analysis of the gag-env deletion junctions of the EAV-HP proviruses from chickens and jungle fowl. (A) Diagram indicating the positions of primers in the gag region (H83REV) and in the env region (H8) flanking the deletion junctions of the 4-kbp EAV-HP provirus. (B) Ethidium bromide-stained PCR products amplified with the H83REV and H8 primers from two layer-type chicken lines, line 0 and brown leghorn (BRL), two meat-type chicken lines, 20 and 21, and two jungle fowl species, RJF and SJF. The EAV-HP1 clone and water were used as positive and negative controls, respectively. PCR products are indicated as I, II, and III, corresponding to the 4-kbp provirus types described in the text.

    Journal: Journal of Virology

    Article Title: Intact EAV-HP Endogenous Retrovirus in Sonnerat's Jungle Fowl

    doi: 10.1128/JVI.75.4.2029-2032.2001

    Figure Lengend Snippet: PCR analysis of the gag-env deletion junctions of the EAV-HP proviruses from chickens and jungle fowl. (A) Diagram indicating the positions of primers in the gag region (H83REV) and in the env region (H8) flanking the deletion junctions of the 4-kbp EAV-HP provirus. (B) Ethidium bromide-stained PCR products amplified with the H83REV and H8 primers from two layer-type chicken lines, line 0 and brown leghorn (BRL), two meat-type chicken lines, 20 and 21, and two jungle fowl species, RJF and SJF. The EAV-HP1 clone and water were used as positive and negative controls, respectively. PCR products are indicated as I, II, and III, corresponding to the 4-kbp provirus types described in the text.

    Article Snippet: PCR was repeated using the Expand high-fidelity PCR system (Boehringer Mannheim) for cloning the 5-kbp SJF EAV-HP PCR product into the pGEM-T vector.

    Techniques: Polymerase Chain Reaction, Staining, Amplification

    Genomic organization of tomato LeFPS genes. A, Thirty micrograms of tomato genomic DNA was digested with the indicated restriction endonuclease and subjected to DNA gel-blot analysis using LeFPS1 cDNA as a hybridization probe. The blot was exposed for 4 d. Numbers on the left indicate size in kb from the markers (1-kb ladder, Gibco BRL, Cleveland). B, Partial map of the chromosomes 10 and 12. The putative location of the two loci (thick line) was deduced from the introgression line showing a polymorphism (dotted lines). LeFPS1 and LeFPS2 were mapped using the 1.3-kb LeFPS1 cDNA insert as a hybridization probe. The 0.18-kb cDNA fragment corresponding to the LeFPS1 3′-UTR was used to locate LeFPS1 on chromosome 12.

    Journal: Plant Physiology

    Article Title: LEFPS1, a Tomato Farnesyl Pyrophosphate Gene Highly Expressed during Early Fruit Development 1

    doi:

    Figure Lengend Snippet: Genomic organization of tomato LeFPS genes. A, Thirty micrograms of tomato genomic DNA was digested with the indicated restriction endonuclease and subjected to DNA gel-blot analysis using LeFPS1 cDNA as a hybridization probe. The blot was exposed for 4 d. Numbers on the left indicate size in kb from the markers (1-kb ladder, Gibco BRL, Cleveland). B, Partial map of the chromosomes 10 and 12. The putative location of the two loci (thick line) was deduced from the introgression line showing a polymorphism (dotted lines). LeFPS1 and LeFPS2 were mapped using the 1.3-kb LeFPS1 cDNA insert as a hybridization probe. The 0.18-kb cDNA fragment corresponding to the LeFPS1 3′-UTR was used to locate LeFPS1 on chromosome 12.

    Article Snippet: A Bam HI site was inserted upstream of the ATG codon in LeFPS1 by PCR using the Expand High fidelity DNA polymerase (Boehringer Mannheim).

    Techniques: Western Blot, Hybridization