Enzymatic polymerisation involving 2鈥?amino-LNA nucleotides

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• 作者：Marie W. Johannsen^a ; Rakesh N. Veedu^a ; ^b ; ^{rakesh@uq.edu.au} ; Andreas Stahl Madsen^a ; Jesper Wengel^a ; ^jwe@sdu.dk
• 关键词：2&prime ; -Amino-LNA ; Locked nucleic acid ; Modified nucleosides ; Primer extension ; Triphosphate
• 刊名：Bioorganic and Medicinal Chemistry Letters
• 出版年：2012
• 期刊代码：10_0960894x
• 类别：ch
• 出版时间：15 May, 2012
• 卷：22
• 期：10
• 页码：3522-3526
• 文件大小：457 K

摘要

The triphosphate of the thymine derivative of 2鈥?amino-LNA (2鈥?amino-LNA-TTP) was synthesised and found to be a good substrate for Phusion庐 HF DNA polymerase, allowing enzymatic synthesis of modified DNA encoded by an unmodified template. To complement this, 2鈥?amino-LNA-T phosphoramidites were incorporated into DNA oligonucleotides which were used as templates for enzymatic synthesis of unmodified DNA using either KOD, KOD XL or Phusion polymerases. 2鈥?Amino-LNA-T in the template and 2鈥?amino-LNA-TTP as a substrate both decreased reaction rate and yield compared to unmodified DNA, especially for sequences with multiple 2鈥?amino-LNA-T nucleotides.