| 摘要 |
<p>Enzymatic synthesis of a repeat region of an oligonucleotide may be performed by the steps of: (a) combining a primer and a blocked nucleotide in the presence of a chain extending enzyme whereby a primer-blocked nucleotide product is formed containing the blocked nucleotide coupled to the primer at its 3'-end; (b) removing the blocking group from the 3'-end of the primer-blocked nucleotide product using a 3'-phosphatase enzyme substantially without removing the 3'-phosphate blocking group from unreacted 3'-phosphate-blocked nucleotide; and (c) repeating the cycle of steps (a) and (b), using the primer-nucleotide product of step (b) as the primer for step (a) in the next cycle, for sufficient cycles to form the oligonucleotide product. These cycles are performed preferably in a single vessel without intermediate purification of oligonucleotide product. Also disclosed is a process for synthesizing an oligonucleotide having a defined sequence including at least one repeat region and one non-repeating region, wherein at least one non-repeating region is synthesized by reaction cycles using the steps of extending a primer with a 3'-blocked nucleotide, inactivating unreacted 3'-blocked nucleotide, and removing the blocking group from the extended primer. The disclosed processes may be used to synthesize repeat regions of oligoribonucleotides.</p> |
