通过T4 RNA连接酶的“模板介导”连接实现寡核糖核苷酸环化：环状锤头状核酶的合成
Oligoribonucleotide circularization by 'template-mediated' ligation with T4 RNA ligase: synthesis of circular hammerhead ribozymes.
作者信息
Wang L, Ruffner D E
机构信息
Department of Bioengineering, University of Utah, 421 Wakara Way, Suite 318, Salt Lake City, UT 84108, USA.
出版信息
Nucleic Acids Res. 1998 May 15;26(10):2502-4. doi: 10.1093/nar/26.10.2502.
Abstract
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.
摘要
环状锤头状核酶是使用T4 RNA连接酶由线性寡核糖核苷酸合成的。一些前体在标准条件下不能有效地环化。对于这些分子，使用DNA模板可使其有效环化。该模板的设计目的是防止前体折叠成不合适的结构。该模板使得在连接反应中长度小至15个核苷酸的环状核酶能够以高达50微摩尔的浓度高效合成。环状产物保留了它们的生物活性。
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