Equimolar addition of oligoribonucleotides with T4 RNA ligase.
T4 induced RNA ligase will join equimolar concentrations of two oligoribonucleotides, (Ap)3C and p(Up) 5, to form a single product, (Ap)3Cp(Up) 5, in high yield. The presence of the 3' phosphate on p(Up)5 prevents the  oligomer from adding to itself. The pH optimum of the reaction is about 7.5, but less of the undesirable adenylated intermediate, App(Up) 5, forms at pH 8.2. The reaction rate is a linear function of oligomer concentration from 3 micronM to 0.6 mM. The data suggest that T4 RNA ligase will be a useful enzyme for the synthesis of oligomers of defined sequence.
D000255:Adenosine Triphosphate / Q000494:pharmacology; D003090:Coliphages / Q000201:enzymology*; D004229:Dithiothreitol / Q000494:pharmacology; D004926:Escherichia coli; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D009154:Mutation; D009841:Oligonucleotides / Q000378:metabolism*; D009843:Oligoribonucleotides / Q000378:metabolism*; D011118:Polynucleotide Ligases / Q000378:metabolism*
D009841:Oligonucleotides; D009843:Oligoribonucleotides; D000255:Adenosine Triphosphate; D011118:Polynucleotide Ligases; D004229:Dithiothreitol