- 1H NMR study of complexation of ethidium bromide with single-stranded noncomplementary desoxytetranucleotide 5′-d(GpApApG) in aqueous solution
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Complexation of the ethidium bromide dye (3,8-diamino-6-phenyl-5-ethylphenanthridine) with single-stranded noncomplementary desoxytetranucleotide 5′-d(GpApApG) in aqueous salt solution was studied by one- and two-dimensional 1H NMR (500 and 600 MHz). The concentration dependences of the proton chemical shifts of the reactant molecules were measured at different temperatures (T1 = 298 K, T2 = 308 K). Investigations of self-association of the tetranucleotide showed that duplices can hardly form in solution. Therefore, dye complexes with single-stranded tetranucleotide play a major role in the equilibrium in solution; this makes it possible to analyze the specifics of interactions of aromatic ligands with single-stranded DNA. Various schemes of complexation are discussed; the equilibrium constants and the limiting values of the proton chemical shifts of ethidium bromide in the complexes are determined. The constants of dye binding to the single-stranded tetranucleotide 5′-d(GAAG) involving only purine bases is approximately an order of magnitude lower than the constants of ethidium bromide complexation with desoxytetranucleotide monomers whose sequences contain alternating types of base in the chain. The relative contents of complexes of different types are analyzed, and peculiarities of dynamic equilibrium, depending on the ratio of concentrations between the dye and the tetranucleotide, are revealed. Based on the data obtained it is concluded that the binding between ethidium bromide and the single-stranded nucleotide is sequence-specific. The estimated values of the induced chemical shifts of the dye protons are used to establish the most probable structures of the 1:1 and 2:1 complexes of ethidium bromide with single-stranded desoxytetranucleotide. 1999 Kluwer Academic/Plenum Publishers.
- Baranovskii,Djimant,Osetrov,Petrenko,Parkes,Davies,Veselkov
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- In vivo light-driven DNA binding and cellular uptake of nucleic acid stains
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Chemical derivatization of nucleic stains such as ethidium bromide or DAPI with tailored, photoresponsive caging groups, allows for on demand spatiotemporal control of their in vivo nucleic acid binding, as well as for improving their cellular uptake. This effect was particularly noteworthy for a nitro-veratryloxycarbonyl-caged derivative of ethidium bromide that, in contrast with the parent stain, is effectively internalized into living cells. The activation strategy works in light-accessible, therapeutically relevant settings, such as human retinas, and can even be applied for the release of active compounds in the eyes of living mice.
- Sanchez, Mateo I.,Martinez-Costas, Jose,Gonzalez, Francisco,Bermudez, Maria A.,Vazquez, M. Eugenio,Mascarenas, Jose L.
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p. 1276 - 1280
(2012/09/08)
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- Improvement of ways to obtain ethidium bromide and synthesis of ethidium ethyl sulfate, a new fluorescent dye for detection of nucleic acids
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Two methods for synthesis of ethidium bromide, a fluorescent dye widely used in molecular biology, were improved. An analog of ethidium bromide, ethidium ethyl sulfate, was synthesized.
- Osadchii,Shubin,Kozlova,Varlamenko,Filipenko,Boyarskikh
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p. 1541 - 1548
(2012/01/14)
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- BIODEGRADABLE POLY(DISULFIDE AMINE)S FOR GENE DELIVERY
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Poly(disulfide amine)s, methods of making, and methods of use are described. Illustrative embodiments of the poly(disulfide amine)s include poly(CBA-DAE), poly(CBA-DAB), and poly(CBA-DAH). These compositions are made by Michael addition between N,N′-cystaminebisacrylamide and N-Boc-protected diamine monomers, followed by N-Boc deprotection. Complexes are formed by mixing the poly(disulfide amine)s with nucleic acid. Delivery of the nucleic acid into cells is carried out by contacting the cells with the nucleic acid/poly(disulfide amine) complexes.
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- DNA-binding peptides searched from the solid-phase combinatorial library with the use of the magnetic beads attaching the target duplex DNA
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We have exhibited successful and rapid screening of DNA-binding peptide ligands from solid-phase library beads with the use of the target DNA- conjugated magnetic beads. The target duplex DNA (3) has a polyether linker between two complementary sequences (T4A3G-ether linker-CT3A4) and is stable in the duplex form during the selection procedure. Finally, 71 pentapeptide sequences were identified from the solid-phase pentapeptide library. From an analysis of the peptide sequences identified in this study, it has been revealed that peptide ligands contain hydrophobic amino acids as the major component. The synthetic peptides with identified sequences and a combination of the major components have exhibited moderate to high binding affinity to the duplex DNA in competition experiments with ethidium-DNA complexes. (C) 2000 Elsevier Science Ltd.
- Rowshon Alam,Maeda, Minoru,Sasaki, Shigeki
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p. 465 - 473
(2007/10/03)
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- Distinctive DNA sequence of E. coli 0157:H7 and its use for the rapid, sensitive and specific detection of 0157:H7 and other enterohemorrhagic E. coli
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The present invention provides isolated nucleic acid sequences corresponding to the hlyA gene, the hlyB gene and the intergenic region between the hlyA gene and the hlyB gene which are present in enterohemorrhagic E. coli. In addition, the present invention provides methods for detecting enterohemorrhagic E. coli by targeting the hlyA gene, the hlyB gene, the intergenic region between the hlyA and the hlyB genes, combinations thereof, or fragments thereof. Such methods rely on nucleic acid probes and amplification primers specific for subsequences of the hlyA gene, the hlyB gene, the intergenic region between the hlyA and the hlyB genes, combination thereof or, fragments thereof. As such, the present provides nucleic acid probes and amplification primers which can be used for the rapid, sensitive and specific amplification and detection of enterohemorrhagic E. coli. In addition, the present invention provides kits embracing the above aspects.
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- SYNTHESE DANS LA CHIMIE DES PHENANTHRIDINES. III. PREPARATION DE QUELQUES MODELES D'ACIDES ω-(DIAMINO-3,8 PHENANTHRIDINYL-6) CARBOXYLIQUES
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New series of ω-(3,8-diamino-6-phenanthridinyl) alkanoic acids are obtained from the acid chloride-esters ROCO(CH2)nCOCl (n =0, 3 to 8; R = Me or Et).Reaction of these with 2-amino-3,8-dicarbethoxyaminobiphenyl leads to the expected 2-acylamino-3,8-dicarbethoxyaminobiphenyls which by cyclization lead to the corresponding methyl or ethyl (3,8-dicarbethoxyamino-6-phenanthridinyl)alkanoates.Quaternarization followed by hydrolysis has been performed to obtain some ω-(3,8-diamino-6-phenanthridinium) alkanoic acid salts.
- Lion, C.,Boukou-Poba, J. P.,Charvy, C.
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p. 171 - 181
(2007/10/02)
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