Modified nucleotides for synthesis of nucleic acids, a kit containing such nucleotides and their use for the production of synthetic nucleic acid sequences or genes

Abstract

A modified nucleotide, intended for the synthesis of long chain nucleic acids by enzymatic processes, comprising a ''natural'' nitrogenous base or a natural nitrogenous base analogue, a ribose or deoxyribose carbohydrate, and at least one phosphate group, characterized in that said nucleotide comprises at least one R group, termed the modifier group, carried by said nitrogenous base or analogue and / or by the oxygen in position 3' of the ribose or deoxyribose molecule, making it possible to block the polymerization of said nucleotide and / or to allow the interaction of said nucleotide with another molecule, such as a protein, during the nucleic acid synthesis, R comprising at least one functional terminal group.

Description

technical field [0001] The present invention falls within the field of synthesis of biologically important functionalized copolymers. More specifically, it relates to nucleotides required for the synthesis of nucleic acids, especially very long nucleic acids, to kits containing these nucleotides, and to their use for the production of synthetic nucleic acid sequences or genes. Background technique [0002] So far, there are two main categories of in vitro nucleic acid synthesis: chemical synthesis and enzymatic synthesis. [0003] The most commonly used method of in vitro chemical synthesis of nucleic acids is the polymerization method using phosphoramidites described by Adams et al. (1983, J. Amer. Chem. Soc. 105:661) and Froehler et al. (1983, Tetrahedron Lett. 24:3171). In this method, each nucleotide to be added is protected on the 5'-OH group in order to prevent uncontrolled polymerization of several nucleotides of the same type. Generally, protection of the 5'-OH gro...

AI Technical Summary

Problems solved by technology

These residues have very deleterious effects on the sequencing process and generally on any use or modification of the nucleic acid

[0021] Regardless of the preserved nucleotide structure, existing modified nucleotides do not meet the expectations of enzymatic synthesis methods

They are characterized by poor use of elongases, positioning of various functional groups, systematic use of modified nitrogenous bases, restriction of retained interactions with complementary nucleotides, numerous deprotection steps and residual scarring. The presence of these nucleotides prevents the use of these nucleotides in the enzymatic synthesis of nucleic acids

[0022] Thus, no satisfactory technical solutions for protected nucleotides compatible with the enzymatic synthesis of nucleic acids, especially for the enzymatic synthesis of very long nucleic acid fragments, have been proposed so far.

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