Poly:desoxy nucleotide(s) prodn. - from 2:desoxy nucleotide 5:tri:phosphate(s) using DNA polymerase fixed to solid phase

Polydesoxy-nucleotides, such as a double helix of alternating 2'-desoxy-adenosine-5'- mono-phosphate, and thymine-5'- mono-phosphate (d(A-T)), are prepd. from 2'-desoxy-nucleotide-5- tri-phosphates using a DNA- polymerase having a free mercapto gp. external to its active centre....

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1. Verfasser: JANOS SAGI, LASZLO OTVOS, ILDIKO FRITSCHE, NEE LUKACS, ANNA SZABOLCS, NEE BORDAS, ATTILA SZEMZO, GABOR KRUPPA ET IVAN PAL
Format: Patent
Sprache:eng ; fre
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Zusammenfassung:Polydesoxy-nucleotides, such as a double helix of alternating 2'-desoxy-adenosine-5'- mono-phosphate, and thymine-5'- mono-phosphate (d(A-T)), are prepd. from 2'-desoxy-nucleotide-5- tri-phosphates using a DNA- polymerase having a free mercapto gp. external to its active centre. This enzyme is fixed by a covalent bond to a solid phase that is capable of linking through this mercapto gp. The prepn. is effected in the presence of divalent cations and opt. a prim. matrix complex, in a suitable buffered soln. at pH 6.5-9.5 and at 15-42 deg.C. The prods. are used as models for a study of the physical, chemical, biochemical and biological properties of nucleic acids. The enzyme can be eliminated at the end of the reaction without losing its activity; it may therefore be used repeatedly and continuously. SYNTHESE DE POLYDESOXYNUCLEOTIDES A PARTIR DE 2-DESOXYNUCLEOTIDES-5-TRIPHOSPHATES A L'AIDE D'UNE ENZYME DNA-POLYMERASE CONTENANT UN GROUPE MERCAPTO-LIBRE EXTERIEUR AU CENTRE ACTIF, EN PRESENCE DE CATIONS DIVALENTS ET EVENTUELLEMENT D'UN COMPLEXE MATRICE-PRIMAIRE DANS UNE SOLUTION AQUEUSE TAMPONNEE A UN PH COMPRIS ENTRE 6,5 ET 9,5 ET ENTRE 15 ET 42C.L'ENZYME UTILISEE EST FIXEE PAR UNE LIAISON COVALENTE A UNE PHASE SOLIDE CAPABLE DE FORMER DES LIAISONS AVEC DES GROUPES MERCAPTO-.