Oligoribonucleotide circularization by 'template-mediated' ligation with T4 RNA ligase: synthesis of circular hammerhead ribozymes.
Circular hammerhead ribozymes were synthesized from linear oligoribonucleotides using T4 RNA ligase. Some of the precursors could not be efficiently circularized under standard conditions. For these molecules, the use of a DNA template allowed their efficient circularization. The template was designed to prevent the precursor from folding into an unsuitable structure. The template allowed circular ribozymes as small as 15 nucleotides in length to be efficiently synthesized at concentrations as high as 50 microM in the ligation reaction. The circular products retained their biological activity.
Wang
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Department of Bioengineering, University of Utah, 421 Wakara Way, Suite 318, Salt Lake City, UT 84108, USA.
Ruffner
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D017122:Bacteriophage T4 / Q000201:enzymology; D004247:DNA; D004270:DNA, Circular / Q000096:biosynthesis* / Q000737:chemistry; D009690:Nucleic Acid Conformation; D009843:Oligoribonucleotides / Q000737:chemistry; D011130:RNA Ligase (ATP); D016337:RNA, Catalytic / Q000096:biosynthesis* / Q000737:chemistry; D013698:Templates, Genetic
D004270:DNA, Circular; D009843:Oligoribonucleotides; D016337:RNA, Catalytic; D004247:DNA; D011130:RNA Ligase (ATP)