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    Wyświetlanie 1-7 z 7
    Tytuł:
    High efficiency method to obtain supercoiled DNA with a commercial plasmid purification kit
    Autorzy:
    Carbone, Antonietta
    Fioretti, Flavia
    Fucci, Laura
    Ausió, Juan
    Piscopo, Marina
    Powiązania:
    https://bibliotekanauki.pl/articles/1039748.pdf
    Data publikacji:
    2012
    Wydawca:
    Polskie Towarzystwo Biochemiczne
    Tematy:
    midi preparation
    supercoiled plasmid isoform
    plasmid purification
    Qiagen kit
    Plasmid DNA
    Opis:
    Supercoiled state corresponds to the active form for plasmid applications. The relaxed circular form of plasmids is often inactive or poorly active. To obtain significant amounts of almost fully supercoiled DNA, we modified the standard protocol of a commercially available Qiagen plasmid purification kit. Our changes led to isolation of almost 100% of the plasmids in the supercoiled state. The modified protocol was used to purify different plasmids with consistent results. The purified plasmids maintain supercoiled state for about two months. The modified protocol is very advantageous because it allows easy DNA production with high degree of supercoiled form at low cost.
    Źródło:
    Acta Biochimica Polonica; 2012, 59, 2; 275-278
    0001-527X
    Pojawia się w:
    Acta Biochimica Polonica
    Dostawca treści:
    Biblioteka Nauki
    Artykuł
    Tytuł:
    The role of actin and microtubule networks in plasmid DNA intracellular trafficking
    Autorzy:
    Ondřej, Vladan
    Lukášová, Emilie
    Falk, Martin
    Kozubek, Stanislav
    Powiązania:
    https://bibliotekanauki.pl/articles/1041059.pdf
    Data publikacji:
    2007
    Wydawca:
    Polskie Towarzystwo Biochemiczne
    Tematy:
    cytoplasmic trafficking
    microtubules
    plasmid DNA-lipid complexes
    actin filaments
    Opis:
    This work is focused on the function of the microtubule and actin networks in plasmid DNA transport during liposomal transfection. We observed strong binding of plasmid DNA-lipid complexes (lipoplexes) to both networks and directional long-range motion of these lipoplexes along the microtubules. Disruption of either of these networks led to the cessation of plasmid transport to the nucleus, a decreased mobility of plasmids, and accumulation of plasmid DNA in large aggregates at the cell periphery. Our findings show an indispensable but different role of both types of cytoskeleton, actin and microtubular, in the processes of gene delivery.
    Źródło:
    Acta Biochimica Polonica; 2007, 54, 3; 657-663
    0001-527X
    Pojawia się w:
    Acta Biochimica Polonica
    Dostawca treści:
    Biblioteka Nauki
    Artykuł
    Tytuł:
    Improved downstream process for the production of plasmid DNA for gene therapy
    Autorzy:
    Urthaler, Jochen
    Buchinger, Wolfgang
    Necina, Roman
    Powiązania:
    https://bibliotekanauki.pl/articles/1041381.pdf
    Data publikacji:
    2005
    Wydawca:
    Polskie Towarzystwo Biochemiczne
    Tematy:
    plasmid DNA
    production
    gene- therapy
    purification
    alkaline lysis
    chromatography
    Opis:
    Gene therapy and genetic vaccines promise to revolutionize the treatment of inherited and acquired diseases. Since viral vectors are generally associated with numerous disadvantages when applied to humans, the administration of naked DNA, or DNA packed into lipo- or polyplexes emerge as viable alternatives. To satisfy the increasing demand for pharmaceutical grade plasmids we developed a novel economic downstream process which overcomes the bottlenecks of common lab-scale techniques and meets all regulatory requirements. After cell lysis by an in-house developed gentle, automated continuous system the sequence of hydrophobic interaction, anion exchange and size exclusion chromatography guarantees the separation of impurities as well as undesired plasmid isoforms. After the consecutive chromatography steps, adjustment of concentration and final filtration are carried out. The final process was proven to be generally applicable and can be used from early clinical phases to market-supply. It is scaleable and free of animal-derived substances, detergents (except lysis) and organic solvents. The process delivers high-purity plasmid DNA of homogeneities up to 98% supercoiled form at a high yield in any desired final buffer.
    Źródło:
    Acta Biochimica Polonica; 2005, 52, 3; 703-711
    0001-527X
    Pojawia się w:
    Acta Biochimica Polonica
    Dostawca treści:
    Biblioteka Nauki
    Artykuł
    Tytuł:
    Intranuclear trafficking of plasmid DNA is mediated by nuclear polymeric proteins lamins and actin
    Autorzy:
    Ondřej, Vladan
    Lukášová, Emilie
    Krejčí, Jana
    Kozubek, Stanislav
    Powiązania:
    https://bibliotekanauki.pl/articles/1040744.pdf
    Data publikacji:
    2008
    Wydawca:
    Polskie Towarzystwo Biochemiczne
    Tematy:
    plasmid DNA
    DNA double-strand breaks
    nuclear actin
    lamin A/C
    Opis:
    Functions of nuclear polymeric proteins such as lamin A/C and actin in transport of plasmid DNA were studied. The results show that the lamina plays an important role in plasmid DNA's entry into the cell nucleus from the cytoplasm. Selective disruption of lamin A/C led to a halt in plasmid DNA transport through the nuclear envelope. Inside the nucleus, plasmid DNA was frequently localized at sites with impaired genome integrity, such as DNA double-strand breaks (DSBs), occurring spontaneously or induced by ionizing radiation. Polymeric actin obviously participates in nuclear transport of plasmid DNA, since inhibition of actin polymerization by latrunculin B disturbed plasmid transport inside the cell nucleus. In addition, precluding of actin polymerization inhibited plasmid co-localization with newly induced DSBs. These findings indicate the crucial role of polymeric actin in intranuclear plasmid transport.
    Źródło:
    Acta Biochimica Polonica; 2008, 55, 2; 307-315
    0001-527X
    Pojawia się w:
    Acta Biochimica Polonica
    Dostawca treści:
    Biblioteka Nauki
    Artykuł
    Tytuł:
    Effect of Mg2+ on kinetics of oxidation of pyrimidines in duplex DNA by potassium permanganate.
    Autorzy:
    Łoziński, Tomasz
    Wierzchowski, Kazimierz
    Powiązania:
    https://bibliotekanauki.pl/articles/1044147.pdf
    Data publikacji:
    2001
    Wydawca:
    Polskie Towarzystwo Biochemiczne
    Tematy:
    oxidation of pyrimidines
    thymine glycol
    magnesium ions
    rate constant of oxidation
    pDS3 plasmid DNA
    quantitative permanganate footprinting
    Opis:
    Potassium permanganate oxidation of pyrimidine bases is often used to probe single-stranded regions in functional DNA-protein complexes. However, so far reactivity of these bases in double-stranded DNA has not been studied quantitatively. We have investigated the kinetics of oxidation of pyrimidines in supercoiled pDS3 plasmid dsDNA by quantitative KMnO4 footprinting, in connection with parallel studies on the effect of Mg2+ on kinetics of oxidation of individual thymines in the single-stranded region of the open transcription complex of Escherichia coli RNA polymerase at a cognate Pa promoter contained in this plasmid. Rate constants of oxidation for pyrimidines, kj, in selected regions of pDS3 DNA, including Pa promoter, were determined under single-hit reaction conditions in the absence and presence of 10 mM MgCl2. Their values appeared to be sequence-dependent and were: (i) the largest for Ts in 5'TA3' and 5'TC3' steps, while 2-4 times smaller for 5'-adjacent ones in TT(A,G,C) and TTT(A) runs, (ii) for Cs in 5'TC3' steps 2-4 fold smaller than for adjacent Ts, and (iii) in the presence of Mg2+ generally larger by a sequence-dependent factor: in 5'TC3' steps of about 2 and 4 for Ts and Cs, respectively, in 5'TA3' steps of TTA and TTTA sequences for 3'-terminal Ts of about 3, while for their 5'-neighbors of a distinctly smaller value of about 2. Comparison of kj data for corresponding Ts located between +1 and -10 regions of Pa promoter in dsDNA and in ssDNA form in the open transcription complex, reported elsewhere, demonstrates that reactivity of pyrimidines in dsDNA is by 2-3 orders of magnitude smaller. The effect of Mg2+ in dsDNA is interpreted in terms of electrostatic barrier to diffusion of MnO4- on DNA surface, which is lowered by diffusive binding of these ions to backbone phosphates, involving also sequence-specific contacts with bases in the minor and major grooves of B-DNA.
    Źródło:
    Acta Biochimica Polonica; 2001, 48, 2; 511-523
    0001-527X
    Pojawia się w:
    Acta Biochimica Polonica
    Dostawca treści:
    Biblioteka Nauki
    Artykuł
    Tytuł:
    Protection of cattle against bovine leukemia virus (BLV) infection could be attained by DNA vaccination
    Autorzy:
    Brillowska, Anna
    Dąbrowski, Sławomir
    Rułka, Jan
    Kubiś, Piotr
    Buzała, Ewa
    Kur, Józef
    Powiązania:
    https://bibliotekanauki.pl/articles/1044457.pdf
    Data publikacji:
    1999
    Wydawca:
    Polskie Towarzystwo Biochemiczne
    Tematy:
    DNA vaccine
    env gene
    expression plasmid
    immunization
    Opis:
    The bovine leukemia virus (BLV) envelope gene encoding extracellular glycoprotein gp51 and transmembrane glycoprotein gp30 was cloned into a vehicle expression vector under the human cytomegalovirus (CMV) intermediate early promoter. The intramuscular injection of this plasmid vector generated a cellular immune response. Seven out of ten cows vaccinated with the DNA construct resisted a drastic challenge (500 BLV-infected lymphocytes as an infectious dose).
    Źródło:
    Acta Biochimica Polonica; 1999, 46, 4; 971-976
    0001-527X
    Pojawia się w:
    Acta Biochimica Polonica
    Dostawca treści:
    Biblioteka Nauki
    Artykuł
    Tytuł:
    1H-Benzimidazole derivatives as mammalian DNA topoisomerase I inhibitors
    Autorzy:
    Alpan, A
    Gunes, H
    Topcu, Zeki
    Powiązania:
    https://bibliotekanauki.pl/articles/1040939.pdf
    Data publikacji:
    2007
    Wydawca:
    Polskie Towarzystwo Biochemiczne
    Tematy:
    1H-benzimidazole derivatives
    type I DNA topoisomerase
    plasmid supercoil relaxation assays
    Opis:
    Benzimidazole is one of the most important heterocyclic groups manifesting various biological properties, such as antibacterial, antifungal, antimicrobial, antiprotozoal and antihelmintic activities. Several benzimidazole derivatives are also active as inhibitors of type I DNA topoisomerases. In this study, three 1H-benzimidazole derivatives with different electronic characteristics at position 5-, namely 5-chloro-4-(1H-benzimidazole-2-yl)phenol (Cpd I), 5-methyl-4-(1H-benzimidazole-2-yl)phenol (Cpd II) and 4-(1H-benzimidazole-2-yl)phenol (Cpd III), were synthesized and evaluated for their effects on mammalian type I DNA topoisomerase activity using quantitative in vitro plasmid supercoil relaxation assays. For the structure elucidation of the compounds, melting points, UV, IR, 1H NMR, 13C NMR, mass spectral data and elemental analyses were interpreted. Among the compounds, 5-methyl-4-(1H-benzimidazole-2-yl)phenol (Cpd II) manifested relatively potent topoisomerase I inhibition.
    Źródło:
    Acta Biochimica Polonica; 2007, 54, 3; 561-565
    0001-527X
    Pojawia się w:
    Acta Biochimica Polonica
    Dostawca treści:
    Biblioteka Nauki
    Artykuł
      Wyświetlanie 1-7 z 7

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