pbluescript sk Stratagene Search Results


  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 93
    Stratagene pbluescript sk stratagene
    Restriction endonuclease map of frxA and rdxA clones from the lambda phage library constructed with genomic DNAs from Mtz-sensitive and -resistant H. pylori strains. The phage clones that carried an frxA gene or an rdxA gene were screened by plaque hybridization. Restriction fragments that contained an frxA or an rdxA gene were identified by Southern hybridization. The restriction fragments that contained the frxA or rdxA gene were inserted into <t>pBluescript</t> SK(+) digested with the same or appropriate restriction enzymes. pGH170 and pGH121 were cloned from the genomic DNA of H. pylori 2600 (Mtz MIC, 2 μg/ml), pGH175 and pGH179 were cloned from the genomic DNA of H. pylori ATCC 700392 (Mtz MIC, 8 μg/ml), pGH174 and pGH101 were cloned from the genomic DNA of H. pylori 6013 (Mtz MIC, 32 μg/ml), pGH178 and pGH160 were cloned from the genomic DNA of H. pylori 1857 (Mtz MIC, 128 μg/ml), and pGH180 and pGH68 were cloned from the genomic DNA of H. pylori 1700 (Mtz MIC, 256 μg/ml). E, Eco RI; E47, Eco 47III; H, Hin dIII; N, Nsi I; S, Sph I; X, Xba I.
    Pbluescript Sk Stratagene, supplied by Stratagene, used in various techniques. Bioz Stars score: 93/100, based on 1553 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pbluescript sk stratagene/product/Stratagene
    Average 93 stars, based on 1553 article reviews
    Price from $9.99 to $1999.99
    pbluescript sk stratagene - by Bioz Stars, 2020-05
    93/100 stars
      Buy from Supplier

    85
    Stratagene pbluescript ii sk stratagene derived plasmids
    Restriction endonuclease map of frxA and rdxA clones from the lambda phage library constructed with genomic DNAs from Mtz-sensitive and -resistant H. pylori strains. The phage clones that carried an frxA gene or an rdxA gene were screened by plaque hybridization. Restriction fragments that contained an frxA or an rdxA gene were identified by Southern hybridization. The restriction fragments that contained the frxA or rdxA gene were inserted into <t>pBluescript</t> SK(+) digested with the same or appropriate restriction enzymes. pGH170 and pGH121 were cloned from the genomic DNA of H. pylori 2600 (Mtz MIC, 2 μg/ml), pGH175 and pGH179 were cloned from the genomic DNA of H. pylori ATCC 700392 (Mtz MIC, 8 μg/ml), pGH174 and pGH101 were cloned from the genomic DNA of H. pylori 6013 (Mtz MIC, 32 μg/ml), pGH178 and pGH160 were cloned from the genomic DNA of H. pylori 1857 (Mtz MIC, 128 μg/ml), and pGH180 and pGH68 were cloned from the genomic DNA of H. pylori 1700 (Mtz MIC, 256 μg/ml). E, Eco RI; E47, Eco 47III; H, Hin dIII; N, Nsi I; S, Sph I; X, Xba I.
    Pbluescript Ii Sk Stratagene Derived Plasmids, supplied by Stratagene, used in various techniques. Bioz Stars score: 85/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pbluescript ii sk stratagene derived plasmids/product/Stratagene
    Average 85 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    pbluescript ii sk stratagene derived plasmids - by Bioz Stars, 2020-05
    85/100 stars
      Buy from Supplier

    90
    Stratagene pbluescript sk pbsk
    Restriction endonuclease map of frxA and rdxA clones from the lambda phage library constructed with genomic DNAs from Mtz-sensitive and -resistant H. pylori strains. The phage clones that carried an frxA gene or an rdxA gene were screened by plaque hybridization. Restriction fragments that contained an frxA or an rdxA gene were identified by Southern hybridization. The restriction fragments that contained the frxA or rdxA gene were inserted into <t>pBluescript</t> SK(+) digested with the same or appropriate restriction enzymes. pGH170 and pGH121 were cloned from the genomic DNA of H. pylori 2600 (Mtz MIC, 2 μg/ml), pGH175 and pGH179 were cloned from the genomic DNA of H. pylori ATCC 700392 (Mtz MIC, 8 μg/ml), pGH174 and pGH101 were cloned from the genomic DNA of H. pylori 6013 (Mtz MIC, 32 μg/ml), pGH178 and pGH160 were cloned from the genomic DNA of H. pylori 1857 (Mtz MIC, 128 μg/ml), and pGH180 and pGH68 were cloned from the genomic DNA of H. pylori 1700 (Mtz MIC, 256 μg/ml). E, Eco RI; E47, Eco 47III; H, Hin dIII; N, Nsi I; S, Sph I; X, Xba I.
    Pbluescript Sk Pbsk, supplied by Stratagene, 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/pbluescript sk pbsk/product/Stratagene
    Average 90 stars, based on 17 article reviews
    Price from $9.99 to $1999.99
    pbluescript sk pbsk - by Bioz Stars, 2020-05
    90/100 stars
      Buy from Supplier

    88
    Stratagene pbluescript sk phagemids
    Restriction endonuclease map of frxA and rdxA clones from the lambda phage library constructed with genomic DNAs from Mtz-sensitive and -resistant H. pylori strains. The phage clones that carried an frxA gene or an rdxA gene were screened by plaque hybridization. Restriction fragments that contained an frxA or an rdxA gene were identified by Southern hybridization. The restriction fragments that contained the frxA or rdxA gene were inserted into <t>pBluescript</t> SK(+) digested with the same or appropriate restriction enzymes. pGH170 and pGH121 were cloned from the genomic DNA of H. pylori 2600 (Mtz MIC, 2 μg/ml), pGH175 and pGH179 were cloned from the genomic DNA of H. pylori ATCC 700392 (Mtz MIC, 8 μg/ml), pGH174 and pGH101 were cloned from the genomic DNA of H. pylori 6013 (Mtz MIC, 32 μg/ml), pGH178 and pGH160 were cloned from the genomic DNA of H. pylori 1857 (Mtz MIC, 128 μg/ml), and pGH180 and pGH68 were cloned from the genomic DNA of H. pylori 1700 (Mtz MIC, 256 μg/ml). E, Eco RI; E47, Eco 47III; H, Hin dIII; N, Nsi I; S, Sph I; X, Xba I.
    Pbluescript Sk Phagemids, supplied by Stratagene, used in various techniques. Bioz Stars score: 88/100, based on 86 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pbluescript sk phagemids/product/Stratagene
    Average 88 stars, based on 86 article reviews
    Price from $9.99 to $1999.99
    pbluescript sk phagemids - by Bioz Stars, 2020-05
    88/100 stars
      Buy from Supplier

    93
    Stratagene pblueskript sk
    Restriction endonuclease map of frxA and rdxA clones from the lambda phage library constructed with genomic DNAs from Mtz-sensitive and -resistant H. pylori strains. The phage clones that carried an frxA gene or an rdxA gene were screened by plaque hybridization. Restriction fragments that contained an frxA or an rdxA gene were identified by Southern hybridization. The restriction fragments that contained the frxA or rdxA gene were inserted into <t>pBluescript</t> SK(+) digested with the same or appropriate restriction enzymes. pGH170 and pGH121 were cloned from the genomic DNA of H. pylori 2600 (Mtz MIC, 2 μg/ml), pGH175 and pGH179 were cloned from the genomic DNA of H. pylori ATCC 700392 (Mtz MIC, 8 μg/ml), pGH174 and pGH101 were cloned from the genomic DNA of H. pylori 6013 (Mtz MIC, 32 μg/ml), pGH178 and pGH160 were cloned from the genomic DNA of H. pylori 1857 (Mtz MIC, 128 μg/ml), and pGH180 and pGH68 were cloned from the genomic DNA of H. pylori 1700 (Mtz MIC, 256 μg/ml). E, Eco RI; E47, Eco 47III; H, Hin dIII; N, Nsi I; S, Sph I; X, Xba I.
    Pblueskript Sk, supplied by Stratagene, used in various techniques. Bioz Stars score: 93/100, based on 37 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pblueskript sk/product/Stratagene
    Average 93 stars, based on 37 article reviews
    Price from $9.99 to $1999.99
    pblueskript sk - by Bioz Stars, 2020-05
    93/100 stars
      Buy from Supplier

    85
    Stratagene pbluescipt sk
    Restriction endonuclease map of frxA and rdxA clones from the lambda phage library constructed with genomic DNAs from Mtz-sensitive and -resistant H. pylori strains. The phage clones that carried an frxA gene or an rdxA gene were screened by plaque hybridization. Restriction fragments that contained an frxA or an rdxA gene were identified by Southern hybridization. The restriction fragments that contained the frxA or rdxA gene were inserted into <t>pBluescript</t> SK(+) digested with the same or appropriate restriction enzymes. pGH170 and pGH121 were cloned from the genomic DNA of H. pylori 2600 (Mtz MIC, 2 μg/ml), pGH175 and pGH179 were cloned from the genomic DNA of H. pylori ATCC 700392 (Mtz MIC, 8 μg/ml), pGH174 and pGH101 were cloned from the genomic DNA of H. pylori 6013 (Mtz MIC, 32 μg/ml), pGH178 and pGH160 were cloned from the genomic DNA of H. pylori 1857 (Mtz MIC, 128 μg/ml), and pGH180 and pGH68 were cloned from the genomic DNA of H. pylori 1700 (Mtz MIC, 256 μg/ml). E, Eco RI; E47, Eco 47III; H, Hin dIII; N, Nsi I; S, Sph I; X, Xba I.
    Pbluescipt Sk, supplied by Stratagene, used in various techniques. Bioz Stars score: 85/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pbluescipt sk/product/Stratagene
    Average 85 stars, based on 10 article reviews
    Price from $9.99 to $1999.99
    pbluescipt sk - by Bioz Stars, 2020-05
    85/100 stars
      Buy from Supplier

    90
    Stratagene pbluescript sk background
    Restriction endonuclease map of frxA and rdxA clones from the lambda phage library constructed with genomic DNAs from Mtz-sensitive and -resistant H. pylori strains. The phage clones that carried an frxA gene or an rdxA gene were screened by plaque hybridization. Restriction fragments that contained an frxA or an rdxA gene were identified by Southern hybridization. The restriction fragments that contained the frxA or rdxA gene were inserted into <t>pBluescript</t> SK(+) digested with the same or appropriate restriction enzymes. pGH170 and pGH121 were cloned from the genomic DNA of H. pylori 2600 (Mtz MIC, 2 μg/ml), pGH175 and pGH179 were cloned from the genomic DNA of H. pylori ATCC 700392 (Mtz MIC, 8 μg/ml), pGH174 and pGH101 were cloned from the genomic DNA of H. pylori 6013 (Mtz MIC, 32 μg/ml), pGH178 and pGH160 were cloned from the genomic DNA of H. pylori 1857 (Mtz MIC, 128 μg/ml), and pGH180 and pGH68 were cloned from the genomic DNA of H. pylori 1700 (Mtz MIC, 256 μg/ml). E, Eco RI; E47, Eco 47III; H, Hin dIII; N, Nsi I; S, Sph I; X, Xba I.
    Pbluescript Sk Background, supplied by Stratagene, used in various techniques. Bioz Stars score: 90/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pbluescript sk background/product/Stratagene
    Average 90 stars, based on 7 article reviews
    Price from $9.99 to $1999.99
    pbluescript sk background - by Bioz Stars, 2020-05
    90/100 stars
      Buy from Supplier

    88
    Stratagene pbluscript sk
    Restriction endonuclease map of frxA and rdxA clones from the lambda phage library constructed with genomic DNAs from Mtz-sensitive and -resistant H. pylori strains. The phage clones that carried an frxA gene or an rdxA gene were screened by plaque hybridization. Restriction fragments that contained an frxA or an rdxA gene were identified by Southern hybridization. The restriction fragments that contained the frxA or rdxA gene were inserted into <t>pBluescript</t> SK(+) digested with the same or appropriate restriction enzymes. pGH170 and pGH121 were cloned from the genomic DNA of H. pylori 2600 (Mtz MIC, 2 μg/ml), pGH175 and pGH179 were cloned from the genomic DNA of H. pylori ATCC 700392 (Mtz MIC, 8 μg/ml), pGH174 and pGH101 were cloned from the genomic DNA of H. pylori 6013 (Mtz MIC, 32 μg/ml), pGH178 and pGH160 were cloned from the genomic DNA of H. pylori 1857 (Mtz MIC, 128 μg/ml), and pGH180 and pGH68 were cloned from the genomic DNA of H. pylori 1700 (Mtz MIC, 256 μg/ml). E, Eco RI; E47, Eco 47III; H, Hin dIII; N, Nsi I; S, Sph I; X, Xba I.
    Pbluscript Sk, supplied by Stratagene, used in various techniques. Bioz Stars score: 88/100, based on 48 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pbluscript sk/product/Stratagene
    Average 88 stars, based on 48 article reviews
    Price from $9.99 to $1999.99
    pbluscript sk - by Bioz Stars, 2020-05
    88/100 stars
      Buy from Supplier

    85
    Stratagene ecorv cut pbluescript sk
    Restriction endonuclease map of frxA and rdxA clones from the lambda phage library constructed with genomic DNAs from Mtz-sensitive and -resistant H. pylori strains. The phage clones that carried an frxA gene or an rdxA gene were screened by plaque hybridization. Restriction fragments that contained an frxA or an rdxA gene were identified by Southern hybridization. The restriction fragments that contained the frxA or rdxA gene were inserted into <t>pBluescript</t> SK(+) digested with the same or appropriate restriction enzymes. pGH170 and pGH121 were cloned from the genomic DNA of H. pylori 2600 (Mtz MIC, 2 μg/ml), pGH175 and pGH179 were cloned from the genomic DNA of H. pylori ATCC 700392 (Mtz MIC, 8 μg/ml), pGH174 and pGH101 were cloned from the genomic DNA of H. pylori 6013 (Mtz MIC, 32 μg/ml), pGH178 and pGH160 were cloned from the genomic DNA of H. pylori 1857 (Mtz MIC, 128 μg/ml), and pGH180 and pGH68 were cloned from the genomic DNA of H. pylori 1700 (Mtz MIC, 256 μg/ml). E, Eco RI; E47, Eco 47III; H, Hin dIII; N, Nsi I; S, Sph I; X, Xba I.
    Ecorv Cut Pbluescript Sk, supplied by Stratagene, 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/ecorv cut pbluescript sk/product/Stratagene
    Average 85 stars, based on 7 article reviews
    Price from $9.99 to $1999.99
    ecorv cut pbluescript sk - by Bioz Stars, 2020-05
    85/100 stars
      Buy from Supplier

    91
    Stratagene pbluescript sk vector
    Functional characterization of rapeseed LPAATs. A, Complementation of an acyltransferase-deficient strain of E. coli . The coding sequences corresponding to the BAT1.13 and BAT1.5 cDNAs were cloned into the <t>pBluescript</t> SK+ vector and transformed into the JC201 plsC − strain. Aliquots from OD 600 = 0.5 cultures were grown in the presence of IPTG. The figure shows the OD after 16 h of growth at various temperatures. JC201 indicates strain only; pBSK indicates strain JC201 transformed with vector only; BAT1.13 and BAT1.5 indicate strain JC201 transformed with the vector containing the coding sequence of BAT1.13 or BAT1.5. Data represent means of three independent transformations. B, Rapeseed microsomal LPAAT activities in E. coli membrane preparations. Membranes were isolated from JC201 cells that had been grown at 30°C after induction by IPTG and were assayed in the presence of saturating concentrations of LPA and [1- 14 C]oleoyl-CoA or [1- 14 C]palmitoyl-CoA, and the quantity of radioactivity in the PA reaction product was integrated. The genotypes of JC201, pBSK, BAT1.13, and BAT1.5 are as in A. The values represent means of three membrane preparations from independent transformant clones.
    Pbluescript Sk Vector, supplied by Stratagene, used in various techniques. Bioz Stars score: 91/100, based on 1142 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pbluescript sk vector/product/Stratagene
    Average 91 stars, based on 1142 article reviews
    Price from $9.99 to $1999.99
    pbluescript sk vector - by Bioz Stars, 2020-05
    91/100 stars
      Buy from Supplier

    85
    Stratagene pbluecript sk
    Functional characterization of rapeseed LPAATs. A, Complementation of an acyltransferase-deficient strain of E. coli . The coding sequences corresponding to the BAT1.13 and BAT1.5 cDNAs were cloned into the <t>pBluescript</t> SK+ vector and transformed into the JC201 plsC − strain. Aliquots from OD 600 = 0.5 cultures were grown in the presence of IPTG. The figure shows the OD after 16 h of growth at various temperatures. JC201 indicates strain only; pBSK indicates strain JC201 transformed with vector only; BAT1.13 and BAT1.5 indicate strain JC201 transformed with the vector containing the coding sequence of BAT1.13 or BAT1.5. Data represent means of three independent transformations. B, Rapeseed microsomal LPAAT activities in E. coli membrane preparations. Membranes were isolated from JC201 cells that had been grown at 30°C after induction by IPTG and were assayed in the presence of saturating concentrations of LPA and [1- 14 C]oleoyl-CoA or [1- 14 C]palmitoyl-CoA, and the quantity of radioactivity in the PA reaction product was integrated. The genotypes of JC201, pBSK, BAT1.13, and BAT1.5 are as in A. The values represent means of three membrane preparations from independent transformant clones.
    Pbluecript Sk, supplied by Stratagene, used in various techniques. Bioz Stars score: 85/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pbluecript sk/product/Stratagene
    Average 85 stars, based on 10 article reviews
    Price from $9.99 to $1999.99
    pbluecript sk - by Bioz Stars, 2020-05
    85/100 stars
      Buy from Supplier

    85
    Stratagene empty pbluescript sk
    Mechanism of increased ch-IAP1 RNA levels in cells expressing v-Rel. DT40 cells were infected with the helper virus REV-A or a retrovirus expressing v-Rel (REV-TW). (A) Nuclear run-on analyses of ch-IAP1 expression. Nuclei isolated from DT40 cells or DT40 cells infected with REV-A or REV-TW were used to produce run-on transcripts in the presence of [ 32 P]UTP. Equivalent amounts of radioactivity from these reactions were hybridized to membranes containing genomic DNA from each cell type, <t>pBluescript</t> plasmid DNA, and plasmids encoding ch-IAP1 and c- jun . Bound radioactivity was visualized by phosphorimager analysis. (B) Half-life analysis of ch-IAP1 RNA in DT40 cells infected with REV-A or REV-TW. Total RNA (10 μg) was isolated from cells treated with actinomycin D (2.5 μg/ml) for 0, 1, 2, 4, 6, 8, and 10 h and analyzed for ch-IAP1 by Northern blot analysis (top panel). Due to low levels of ch-IAP1 mRNA expression in REV-A-infected DT40 cells, a longer exposure of the Northern blot from these cells is shown than for REV-TW-infected cells. ch-IAP1 RNA expression was normalized for loading by comparing the expression of β-actin (lower panel).
    Empty Pbluescript Sk, supplied by Stratagene, 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/empty pbluescript sk/product/Stratagene
    Average 85 stars, based on 12 article reviews
    Price from $9.99 to $1999.99
    empty pbluescript sk - by Bioz Stars, 2020-05
    85/100 stars
      Buy from Supplier

    Image Search Results


    Restriction endonuclease map of frxA and rdxA clones from the lambda phage library constructed with genomic DNAs from Mtz-sensitive and -resistant H. pylori strains. The phage clones that carried an frxA gene or an rdxA gene were screened by plaque hybridization. Restriction fragments that contained an frxA or an rdxA gene were identified by Southern hybridization. The restriction fragments that contained the frxA or rdxA gene were inserted into pBluescript SK(+) digested with the same or appropriate restriction enzymes. pGH170 and pGH121 were cloned from the genomic DNA of H. pylori 2600 (Mtz MIC, 2 μg/ml), pGH175 and pGH179 were cloned from the genomic DNA of H. pylori ATCC 700392 (Mtz MIC, 8 μg/ml), pGH174 and pGH101 were cloned from the genomic DNA of H. pylori 6013 (Mtz MIC, 32 μg/ml), pGH178 and pGH160 were cloned from the genomic DNA of H. pylori 1857 (Mtz MIC, 128 μg/ml), and pGH180 and pGH68 were cloned from the genomic DNA of H. pylori 1700 (Mtz MIC, 256 μg/ml). E, Eco RI; E47, Eco 47III; H, Hin dIII; N, Nsi I; S, Sph I; X, Xba I.

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: Analysis of rdxA and Involvement of Additional Genes Encoding NAD(P)H Flavin Oxidoreductase (FrxA) and Ferredoxin-Like Protein (FdxB) in Metronidazole Resistance of Helicobacter pylori

    doi:

    Figure Lengend Snippet: Restriction endonuclease map of frxA and rdxA clones from the lambda phage library constructed with genomic DNAs from Mtz-sensitive and -resistant H. pylori strains. The phage clones that carried an frxA gene or an rdxA gene were screened by plaque hybridization. Restriction fragments that contained an frxA or an rdxA gene were identified by Southern hybridization. The restriction fragments that contained the frxA or rdxA gene were inserted into pBluescript SK(+) digested with the same or appropriate restriction enzymes. pGH170 and pGH121 were cloned from the genomic DNA of H. pylori 2600 (Mtz MIC, 2 μg/ml), pGH175 and pGH179 were cloned from the genomic DNA of H. pylori ATCC 700392 (Mtz MIC, 8 μg/ml), pGH174 and pGH101 were cloned from the genomic DNA of H. pylori 6013 (Mtz MIC, 32 μg/ml), pGH178 and pGH160 were cloned from the genomic DNA of H. pylori 1857 (Mtz MIC, 128 μg/ml), and pGH180 and pGH68 were cloned from the genomic DNA of H. pylori 1700 (Mtz MIC, 256 μg/ml). E, Eco RI; E47, Eco 47III; H, Hin dIII; N, Nsi I; S, Sph I; X, Xba I.

    Article Snippet: The appropriate restriction fragments from the screened phage clones carrying the frxA and rdxA genes were identified by hybridization with the same probes and inserted into pBluescript SK(+) or H. pylori-E. coli shuttle vector pHel2 ( ).

    Techniques: Clone Assay, Construct, Hybridization

    DNase I footprinting analyses of protein-DNA interactions on the DS fragment. A , pKS-AHF contains a 251-bp sequence (HinfI site at 8945 nt; AluI site at 9195 nt of the EBV B95-8 genome) carrying DS on the pBluescript vector. B , a 0.44-kbp DNA of pKS-AHF

    Journal: The Journal of Biological Chemistry

    Article Title: Epstein-Barr Nuclear Antigen 1 (EBNA1)-dependent Recruitment of Origin Recognition Complex (Orc) on oriP of Epstein-Barr Virus with Purified Proteins

    doi: 10.1074/jbc.M112.368456

    Figure Lengend Snippet: DNase I footprinting analyses of protein-DNA interactions on the DS fragment. A , pKS-AHF contains a 251-bp sequence (HinfI site at 8945 nt; AluI site at 9195 nt of the EBV B95-8 genome) carrying DS on the pBluescript vector. B , a 0.44-kbp DNA of pKS-AHF

    Article Snippet: An oriP plasmid, pKS-EX, and its deletion derivatives, pKS-EXΔDS and pKS-DS, as well as EBNA1-encoding DNA (derived from the EBV strain B95-8) were kindly provided by Dr. Shirakata ( ). pKS-ARV, a DS-containing plasmid, was created by inserting an EcoRV-AluI fragment (8995–9195 nt of EBV B95-8) into a blunted SalI site of pBluescript II KS(−) vector (Stratagene). pKS-AHF, another DS-containing plasmid, was created by inserting a blunted HinfI-AluI fragment (8945–9195 nt of EBV B95-8) between SmaI and blunted SalI sites of pBluescript II KS(−). pSOP-T48 was made by replacing the bla-Lac Z′ (2126–2657 nt) portion of pBluescript II SK(−) with the SV-neo (G418R ) unit from pMAMneo (Stratagene), followed by inserting 48x tet O ( ) between SalI and XhoI sites and full-length oriP (from pKS-EX) between EcoRI and XbaI sites of its multicloning site.

    Techniques: Footprinting, Sequencing, Plasmid Preparation

    SATB1 binds to genomic DNA in vivo . ( A ) PCR amplification of immunoprecipitated DNA. After purification of immunoprecipitated DNA and linker ligation, PCR amplification was performed as described in the Materials and Methods section using 17 cycles. PCR products from preimmune (lane 1 ) and anti-SATB1 immunoprecipitated DNA (lane 2 ) were examined by 1% agarose gel electrophoresis followed by staining with ethidium bromide. Lane M , 1-kb molecular size markers. ( B ) Gel mobility shift with anti-SATB1 immunoprecipitated DNA. PCR products shown by a bracket in A were isolated from the gel and cloned into pBluescript. Insert DNA in different size ranges was tested for SATB1-binding activity. DNA in the range of 0.6 kb (lanes 1–3 ) and 1-kb ranges (lanes 4–6 ) are shown. Lanes 1 and 4 contain 0 nM; lanes 2 and 5 , 8 nM; lanes 3 and 6 , 16 nM GST– SATB1 protein. ( C ) PCR amplification of preimmune immunoprecipitated DNA. PCR products were generated using 35 cycles of amplification, and the resulting DNA was visualized in 1% agarose gel stained with ethidium bromide (lane 1 ). The region shown by a bracket was cloned into pBluescript. Lane M , 1-kb molecular size markers. ( D ) Gel mobility shift assay with preimmune immunoprecipitated DNA. Inserts derived from DNA shown by the bracket from C were isolated and used in a gel mobility shift assay with GST–SATB1 protein. Lanes 1–3 contain 0, 8, and 16 nM GST–SATB1 protein, respectively. ( E ) Gel mobility shift assay with precleared anti-SATB1 antibody. A similar experiment as described for C and D was performed with precleared anti-SATB1 antiserum, which was prepared by adding SATB1 to the serum, followed by immunoprecipitation. After a 35-cycle amplification, the cloned DNA failed to demonstrate a gel shift with GST–SATB1. Lanes 1–3 contain 0, 8, and 16 nM GST–SATB1, respectively.

    Journal: The Journal of Cell Biology

    Article Title: The Genomic Sequences Bound to Special AT-rich Sequence-binding Protein 1 (SATB1) In Vivo in Jurkat T Cells Are Tightly Associated with the Nuclear Matrix at the Bases of the Chromatin Loops

    doi:

    Figure Lengend Snippet: SATB1 binds to genomic DNA in vivo . ( A ) PCR amplification of immunoprecipitated DNA. After purification of immunoprecipitated DNA and linker ligation, PCR amplification was performed as described in the Materials and Methods section using 17 cycles. PCR products from preimmune (lane 1 ) and anti-SATB1 immunoprecipitated DNA (lane 2 ) were examined by 1% agarose gel electrophoresis followed by staining with ethidium bromide. Lane M , 1-kb molecular size markers. ( B ) Gel mobility shift with anti-SATB1 immunoprecipitated DNA. PCR products shown by a bracket in A were isolated from the gel and cloned into pBluescript. Insert DNA in different size ranges was tested for SATB1-binding activity. DNA in the range of 0.6 kb (lanes 1–3 ) and 1-kb ranges (lanes 4–6 ) are shown. Lanes 1 and 4 contain 0 nM; lanes 2 and 5 , 8 nM; lanes 3 and 6 , 16 nM GST– SATB1 protein. ( C ) PCR amplification of preimmune immunoprecipitated DNA. PCR products were generated using 35 cycles of amplification, and the resulting DNA was visualized in 1% agarose gel stained with ethidium bromide (lane 1 ). The region shown by a bracket was cloned into pBluescript. Lane M , 1-kb molecular size markers. ( D ) Gel mobility shift assay with preimmune immunoprecipitated DNA. Inserts derived from DNA shown by the bracket from C were isolated and used in a gel mobility shift assay with GST–SATB1 protein. Lanes 1–3 contain 0, 8, and 16 nM GST–SATB1 protein, respectively. ( E ) Gel mobility shift assay with precleared anti-SATB1 antibody. A similar experiment as described for C and D was performed with precleared anti-SATB1 antiserum, which was prepared by adding SATB1 to the serum, followed by immunoprecipitation. After a 35-cycle amplification, the cloned DNA failed to demonstrate a gel shift with GST–SATB1. Lanes 1–3 contain 0, 8, and 16 nM GST–SATB1, respectively.

    Article Snippet: Amplification was performed using both KS and SK primers from pBluescript and Pfu polymerase (Stratagene).

    Techniques: In Vivo, Polymerase Chain Reaction, Amplification, Immunoprecipitation, Purification, Ligation, Agarose Gel Electrophoresis, Staining, Mobility Shift, Isolation, Clone Assay, Binding Assay, Activity Assay, Generated, Derivative Assay, Electrophoretic Mobility Shift Assay

    Schematic representations of PAX6 and PAX6(5a) and plasmids used as internal standards. A : Structure of PAX6, 422 amino acids and PAX6(5a), 436 amino acids. Exon 5a encodes a 14 amino acid insertion into the PAI subdomain of the paired domain disrupting its ability to bind DNA. RED subdomain and homeodomain (HD) are also shown. B : Internal standards (mimics) were created by subcloning three copies of a synthetic oligonucleotide into a unique Pst I site of the PAX6 and PAX6(5a) PCR generated fragments in pBluescript SK II as described in the Materials and Methods. The sizes of PCR products for PAX6, mimic PAX6, PAX6(5a) and mimic PAX6(5a) are 289, 431, 331, and 481 bases, respectively.

    Journal: Molecular vision

    Article Title: Quantitation of PAX6 and PAX6(5a) transcript levels in adult human lens, cornea, and monkey retina

    doi:

    Figure Lengend Snippet: Schematic representations of PAX6 and PAX6(5a) and plasmids used as internal standards. A : Structure of PAX6, 422 amino acids and PAX6(5a), 436 amino acids. Exon 5a encodes a 14 amino acid insertion into the PAI subdomain of the paired domain disrupting its ability to bind DNA. RED subdomain and homeodomain (HD) are also shown. B : Internal standards (mimics) were created by subcloning three copies of a synthetic oligonucleotide into a unique Pst I site of the PAX6 and PAX6(5a) PCR generated fragments in pBluescript SK II as described in the Materials and Methods. The sizes of PCR products for PAX6, mimic PAX6, PAX6(5a) and mimic PAX6(5a) are 289, 431, 331, and 481 bases, respectively.

    Article Snippet: Oligonucleotides used for PAX6/PAX6(5a) (AGC TAT CGA TGG CAG AAG ATT GTA GAG and CGG GAT CCG ATG ACA CGC TTG GTA TG) and TBP (CGG GAT CCT TCC ACT CAC AGA CTC TCA C and CGG AAT TCG CTC TCT TAT CCT CAT GAT TAC C) amplification, were extended with non-complementary sequences (blue) with restriction sites ClaI, BamHI , and EcoRI (red) and subcloned as a ClaI-BamHI (PAX6) and a BamHI-EcoRI (TBP) fragments into a pBluescript SK II (Stratagene, LaJolla, CA).

    Techniques: Subcloning, Polymerase Chain Reaction, Generated

    Functional characterization of rapeseed LPAATs. A, Complementation of an acyltransferase-deficient strain of E. coli . The coding sequences corresponding to the BAT1.13 and BAT1.5 cDNAs were cloned into the pBluescript SK+ vector and transformed into the JC201 plsC − strain. Aliquots from OD 600 = 0.5 cultures were grown in the presence of IPTG. The figure shows the OD after 16 h of growth at various temperatures. JC201 indicates strain only; pBSK indicates strain JC201 transformed with vector only; BAT1.13 and BAT1.5 indicate strain JC201 transformed with the vector containing the coding sequence of BAT1.13 or BAT1.5. Data represent means of three independent transformations. B, Rapeseed microsomal LPAAT activities in E. coli membrane preparations. Membranes were isolated from JC201 cells that had been grown at 30°C after induction by IPTG and were assayed in the presence of saturating concentrations of LPA and [1- 14 C]oleoyl-CoA or [1- 14 C]palmitoyl-CoA, and the quantity of radioactivity in the PA reaction product was integrated. The genotypes of JC201, pBSK, BAT1.13, and BAT1.5 are as in A. The values represent means of three membrane preparations from independent transformant clones.

    Journal: Plant Physiology

    Article Title: Expression of Rapeseed Microsomal Lysophosphatidic Acid Acyltransferase Isozymes Enhances Seed Oil Content in Arabidopsis 1

    doi: 10.1104/pp.109.148247

    Figure Lengend Snippet: Functional characterization of rapeseed LPAATs. A, Complementation of an acyltransferase-deficient strain of E. coli . The coding sequences corresponding to the BAT1.13 and BAT1.5 cDNAs were cloned into the pBluescript SK+ vector and transformed into the JC201 plsC − strain. Aliquots from OD 600 = 0.5 cultures were grown in the presence of IPTG. The figure shows the OD after 16 h of growth at various temperatures. JC201 indicates strain only; pBSK indicates strain JC201 transformed with vector only; BAT1.13 and BAT1.5 indicate strain JC201 transformed with the vector containing the coding sequence of BAT1.13 or BAT1.5. Data represent means of three independent transformations. B, Rapeseed microsomal LPAAT activities in E. coli membrane preparations. Membranes were isolated from JC201 cells that had been grown at 30°C after induction by IPTG and were assayed in the presence of saturating concentrations of LPA and [1- 14 C]oleoyl-CoA or [1- 14 C]palmitoyl-CoA, and the quantity of radioactivity in the PA reaction product was integrated. The genotypes of JC201, pBSK, BAT1.13, and BAT1.5 are as in A. The values represent means of three membrane preparations from independent transformant clones.

    Article Snippet: JC201 cells were transformed with the coding sequences corresponding to the BAT1.13 or BAT1.5 cDNA cloned into the pBluescript SK+ vector (Stratagene).

    Techniques: Functional Assay, Clone Assay, Plasmid Preparation, Transformation Assay, Sequencing, Isolation, Radioactivity

    Mechanism of increased ch-IAP1 RNA levels in cells expressing v-Rel. DT40 cells were infected with the helper virus REV-A or a retrovirus expressing v-Rel (REV-TW). (A) Nuclear run-on analyses of ch-IAP1 expression. Nuclei isolated from DT40 cells or DT40 cells infected with REV-A or REV-TW were used to produce run-on transcripts in the presence of [ 32 P]UTP. Equivalent amounts of radioactivity from these reactions were hybridized to membranes containing genomic DNA from each cell type, pBluescript plasmid DNA, and plasmids encoding ch-IAP1 and c- jun . Bound radioactivity was visualized by phosphorimager analysis. (B) Half-life analysis of ch-IAP1 RNA in DT40 cells infected with REV-A or REV-TW. Total RNA (10 μg) was isolated from cells treated with actinomycin D (2.5 μg/ml) for 0, 1, 2, 4, 6, 8, and 10 h and analyzed for ch-IAP1 by Northern blot analysis (top panel). Due to low levels of ch-IAP1 mRNA expression in REV-A-infected DT40 cells, a longer exposure of the Northern blot from these cells is shown than for REV-TW-infected cells. ch-IAP1 RNA expression was normalized for loading by comparing the expression of β-actin (lower panel).

    Journal: Journal of Virology

    Article Title: Differential Regulation of the Inhibitor of Apoptosis ch-IAP1 by v-rel and the Proto-Oncogene c-rel

    doi: 10.1128/JVI.76.23.11960-11970.2002

    Figure Lengend Snippet: Mechanism of increased ch-IAP1 RNA levels in cells expressing v-Rel. DT40 cells were infected with the helper virus REV-A or a retrovirus expressing v-Rel (REV-TW). (A) Nuclear run-on analyses of ch-IAP1 expression. Nuclei isolated from DT40 cells or DT40 cells infected with REV-A or REV-TW were used to produce run-on transcripts in the presence of [ 32 P]UTP. Equivalent amounts of radioactivity from these reactions were hybridized to membranes containing genomic DNA from each cell type, pBluescript plasmid DNA, and plasmids encoding ch-IAP1 and c- jun . Bound radioactivity was visualized by phosphorimager analysis. (B) Half-life analysis of ch-IAP1 RNA in DT40 cells infected with REV-A or REV-TW. Total RNA (10 μg) was isolated from cells treated with actinomycin D (2.5 μg/ml) for 0, 1, 2, 4, 6, 8, and 10 h and analyzed for ch-IAP1 by Northern blot analysis (top panel). Due to low levels of ch-IAP1 mRNA expression in REV-A-infected DT40 cells, a longer exposure of the Northern blot from these cells is shown than for REV-TW-infected cells. ch-IAP1 RNA expression was normalized for loading by comparing the expression of β-actin (lower panel).

    Article Snippet: Briefly, genomic DNA (2 μg) isolated from each cell line and linearized plasmids (1 μg) encoding full-length c- jun or ch-IAP1 or empty pBluescript SK(−) (Stratagene, La Jolla, Calif.) were denatured, neutralized, and slot blotted onto Hybond N+ (Amersham Pharmacia Biotech, Piscataway, N.J.).

    Techniques: Expressing, Infection, Isolation, Radioactivity, Plasmid Preparation, Northern Blot, RNA Expression