PREPARATION OF OLIGORIBONUCLEOTIDES
First Claim
1. A PROCESS FOR THE PREPARATION OF OLIGORIBONUCLEOTIDES OF DEFINED AND PREDETERMINED SEQUENCE CONTAINING THE NATURAL 3'"'"''"'"'-5'"'"''"'"'-INNTERNUCLEOTIDE LINKAGE, WHICH COMPRISES:
- (A) REACTING DESIRED OLIGONUCLEOTIDE WITH A 2'"'"''"'"'(3'"'"''"'"')-O-ACYL NUCLEOSIDE DIPHOSPHATE IN THE PRESENCE OF POLYNUCLEOTIDE PHOSPHORYLASE, (B) ISOLATING THE PRODUCT FROM THE REACTION MIXTURE, AND (C) REMOVING THE 2'"'"''"'"'(3'"'"''"'"')-O-ACYL BLOCKING GROUP.
0 Assignments
0 Petitions
Accused Products
Abstract
A process for the preparation of oligoribonucleotides of definend predetermined sequence containing the natural 3'"'"''"'"'-5'"'"''"'"'internucleotide linkage, which comprises reacting an oligonucleotide with a monofunctional substrate in the presence of polynucleotide phosphorylase, to form a product which is longer, by one monomeric unit than the initial oligonucleotide, isolating the product from the reaction mixture, removing the blocking agent and repeating the process a desired number of times, adding by each step a desired unit until the desired product of predetermined length and sequence is obtained.
-
Citations
6 Claims
-
1. A PROCESS FOR THE PREPARATION OF OLIGORIBONUCLEOTIDES OF DEFINED AND PREDETERMINED SEQUENCE CONTAINING THE NATURAL 3'"'"''"'"'-5'"'"''"'"'-INNTERNUCLEOTIDE LINKAGE, WHICH COMPRISES:
- (A) REACTING DESIRED OLIGONUCLEOTIDE WITH A 2'"'"''"'"'(3'"'"''"'"')-O-ACYL NUCLEOSIDE DIPHOSPHATE IN THE PRESENCE OF POLYNUCLEOTIDE PHOSPHORYLASE, (B) ISOLATING THE PRODUCT FROM THE REACTION MIXTURE, AND (C) REMOVING THE 2'"'"''"'"'(3'"'"''"'"')-O-ACYL BLOCKING GROUP.
-
2. A process as claimed in claim 1, characterized in that the polynucleotide phosphorylase is used in watersoluble form.
-
3. A process as claimed in claim 1, characterized in that the polynucleotide phosphorylase is used in carrier-bound insoluble form.
-
4. A process as claimed in claim 1, wherein the 2'"'"''"'"'(3'"'"''"'"')-O-acylnucleoside diphosphate is a 2'"'"''"'"'(3'"'"''"'"')-O-acyl ester of ADP, GDP, UDP, CDP, IDP, XDP.
-
5. A process as claimed in claim 1, wherein the 2'"'"''"'"' 3'"'"''"'"'(-O-acyl blocking group is a propionyl, butyryl, valeryl, isobutyryl, isovaleryl or methoxybenzoyl group.
-
6. A process as claimed in claim 1, wherein the reaction, isolation and removal steps (a), (b) and (c) are repeated, in sequence, utilizing as the initial reactant the product isolated in the aforesaid step (b), to produce an oligoribonucleotide of any predetermined length and sequence.
Specification