Enzymatic synthesis of oligonucleotides of defined sequence. Addition of short blocks of nucleotide residues to oligonucleotide primers. 1975

S Gillam, and K Waterman, and M Smith

Polynucleotide phosphorylase from Escherichia coli can be used to catalyse the addition of short tracts of deoxyadenylate residues to the 3'-termini of deoxyribooligonucleotides of the type pdAn-dN (where dN = dC, dT or dG) using dADP as donor. Similarly, the enzyme can also be used to catalyse the addition of short tracts of adenylate residues to the 3'-termini of ribooligonucleotides of the type An-N (where N = C, U or G) using ADP as donor. In the ribooligonucleotide series, phosphorolytic cleavage of the primer oligonucleotides is significant and results in the concommitant production of oligoadenylates lacking the N residue. Oligomers of the same length, with and without the residue N, were readily separated by thermal elution from cellulose-pdT9 columns. This latter procedure therefore provides a simple method for purification of the oligoadenylates containing an internal base substitution and it also provides a convenient assay for oligonucleotide phosphorolysis.

UI MeSH Term Description Entries
D009841 Oligonucleotides Polymers made up of a few (2-20) nucleotides. In molecular genetics, they refer to a short sequence synthesized to match a region where a mutation is known to occur, and then used as a probe (OLIGONUCLEOTIDE PROBES). (Dorland, 28th ed) Oligonucleotide
D010727 Phosphoric Diester Hydrolases A class of enzymes that catalyze the hydrolysis of one of the two ester bonds in a phosphodiester compound. EC 3.1.4. Phosphodiesterase,Phosphodiesterases,Hydrolases, Phosphoric Diester
D011063 Poly A-U A double-stranded polyribonucleotide comprising polyadenylic and polyuridylic acids. Polyadenylic-Polyuridylic Acid,Poly r(A-U),Poly(rA)-Poly(rU),Acid, Polyadenylic-Polyuridylic,Poly A U,Polyadenylic Polyuridylic Acid
D011117 Polyribonucleotide Nucleotidyltransferase An enzyme of the transferase class that catalyzes the reaction RNA(n+1) and orthophosphate to yield RNA(n) and a nucleoside diphosphate, or the reverse reaction. ADP, IDP, GDP, UDP, and CDP can act as donors in the latter case. (From Dorland, 27th ed) EC 2.7.7.8. Polynucleotide Phosphorylase,Nucleotidyltransferase, Polyribonucleotide,Phosphorylase, Polynucleotide
D011119 Polynucleotides BIOPOLYMERS composed of NUCLEOTIDES covalently bonded in a chain. The most common examples are DNA and RNA chains. Polynucleotide
D003854 Deoxyribonucleotides A purine or pyrimidine base bonded to a DEOXYRIBOSE containing a bond to a phosphate group. Deoxyribonucleotide
D004926 Escherichia coli A species of gram-negative, facultatively anaerobic, rod-shaped bacteria (GRAM-NEGATIVE FACULTATIVELY ANAEROBIC RODS) commonly found in the lower part of the intestine of warm-blooded animals. It is usually nonpathogenic, but some strains are known to produce DIARRHEA and pyogenic infections. Pathogenic strains (virotypes) are classified by their specific pathogenic mechanisms such as toxins (ENTEROTOXIGENIC ESCHERICHIA COLI), etc. Alkalescens-Dispar Group,Bacillus coli,Bacterium coli,Bacterium coli commune,Diffusely Adherent Escherichia coli,E coli,EAggEC,Enteroaggregative Escherichia coli,Enterococcus coli,Diffusely Adherent E. coli,Enteroaggregative E. coli,Enteroinvasive E. coli,Enteroinvasive Escherichia coli
D006163 Ribonuclease T1 An enzyme catalyzing the endonucleolytic cleavage of RNA at the 3'-position of a guanylate residue. EC 3.1.27.3. Guanyloribonuclease,RNase T1,Ribonuclease N1,Aspergillus oryzae Ribonuclease,Guanyl-Specific RNase,RNase Apl,RNase F1,RNase Pch 1,RNase ST,Ribonuclease F1,Ribonuclease F2,Ribonuclease ST,Ribonuclease T-1,T 1 RNase,Guanyl Specific RNase,RNase, Guanyl-Specific,RNase, T 1,Ribonuclease T 1,Ribonuclease, Aspergillus oryzae
D000227 Adenine Nucleotides Adenine Nucleotide,Adenosine Phosphate,Adenosine Phosphates,Nucleotide, Adenine,Nucleotides, Adenine,Phosphate, Adenosine,Phosphates, Adenosine
D001483 Base Sequence The sequence of PURINES and PYRIMIDINES in nucleic acids and polynucleotides. It is also called nucleotide sequence. DNA Sequence,Nucleotide Sequence,RNA Sequence,DNA Sequences,Base Sequences,Nucleotide Sequences,RNA Sequences,Sequence, Base,Sequence, DNA,Sequence, Nucleotide,Sequence, RNA,Sequences, Base,Sequences, DNA,Sequences, Nucleotide,Sequences, RNA

Related Publications

S Gillam, and K Waterman, and M Smith
August 1972, Nature: New biology,
S Gillam, and K Waterman, and M Smith
September 1972, Biochemical and biophysical research communications,
S Gillam, and K Waterman, and M Smith
December 1974, Nucleic acids research,
S Gillam, and K Waterman, and M Smith
November 1980, Nucleic acids research,
S Gillam, and K Waterman, and M Smith
April 1978, The Journal of biological chemistry,
S Gillam, and K Waterman, and M Smith
October 1987, DNA (Mary Ann Liebert, Inc.),
S Gillam, and K Waterman, and M Smith
January 1977, Proceedings of the National Academy of Sciences of the United States of America,
Copied contents to your clipboard!