Compositions for enzyme catalyzed template-independent production of phosphodiester bonds using protected nucleotides

摘要

The starting substrate and 3''-hydroxyl, the nucleoside 5''-triphosphate is selected in the order of sequence as planned; provides a starting substrate comprising a nucleotide with (57) unprotected 3''-hydroxyl groups and the summary of the free and due to the fact that the base, and 5 between the unprotected 3''-hydroxyl groups of the starting substrate mononucleotides and 5''-phosphate ''to 3'' to allow binding solution containing the enzyme catalytic amount capable of catalyzing phosphodiester bonds, expected A method for producing stepwise phosphodiester bond results in the synthesis of a polynucleotide sequences, the nucleotides between desired, mononucleotides have a protective 3''-hydroxyl group, protected mononucleotides shared initiating substrate The bind with, added further inhibited 3''-protecting group. Drawing is a schematic of the enzymatic synthesis of oligonucleotides under prevent the nucleoside 5''-triphosphate having a protective group at the 3 ''position and template-dependent polymerase. It is also described the addition to the free polynucleotide chain mononucleotide which is immobilized algae solid support.