PMID: 7541130Jun 11, 1995Paper

Enzymatic synthesis of 2'-modified nucleic acids: identification of important phosphate and ribose moieties in RNase P substrates

Nucleic Acids Research
F ConradG Krupp

Abstract

For the first time mosaic nucleic acids composed of 50% RNA and 50% DNA can be obtained as transcripts with T7 RNA polymerase. Two NTPs could be replaced simultaneously in a transcription reaction. This means more than 40 deoxynucleotides were inserted in one transcript. Previously, a maximum of two deoxynucleotides could be incorporated and 2'-O-methyl-NTPs were not substrates at all. We obtained reasonable transcript yields with a maximal level of 99% 2'-O-methyl-NTPs, and the products contained up to 58% 2'-O-methylnucleotides at more than 20 positions. Sequence-specific nucleotide incorporation was monitored by sequence ladders (partial alkali or iodine cleavage). No base misincorporations were detected with 100% dGTP, dCTP and dTTP, and with partial incorporation of dATP alpha S, 2'-O-methyl-GTP alpha S and 2'-O-methyl-CTP alpha S, whereas they were found with dATP, 2'-O-methyl-ATP alpha S and 2'-O-methyl-UTP alpha S. Quantitative data allow predetermined modification levels of partially modified transcripts. Highly modified transcripts can be used for structural and functional studies, in modification interference approaches and for in vitro evolution procedures. Modification interference studies revealed a small number o...Continue Reading

References

Feb 25, 1991·Nucleic Acids Research·D L ThurlowT L Marsh
Jul 15, 1991·Proceedings of the National Academy of Sciences of the United States of America·D SchatzF Eckstein
Jun 1, 1989·Proceedings of the National Academy of Sciences of the United States of America·S Tabor, C C Richardson
Jan 26, 1988·Biochemistry·W S el-DeiryK M Downey
Dec 1, 1973·Proceedings of the National Academy of Sciences of the United States of America·J D Karkas
Nov 25, 1994·Nucleic Acids Research·H AurupF Eckstein

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Citations

Dec 10, 1998·Biopolymers·S A Strobel
Jan 1, 1995·Molecular Biology Reports·G Krupp
Jul 24, 2003·Journal of Molecular Biology·Ivelitza Garcia, Kevin M Weeks
Apr 1, 1997·Current Opinion in Biotechnology·S W May
Dec 10, 1998·Nature Structural Biology·A A SzewczakS A Strobel
Aug 10, 2011·Proceedings of the National Academy of Sciences of the United States of America·Simon G TrevinoJack W Szostak
Apr 7, 1999·Antisense & Nucleic Acid Drug Development·M WernerS T George
Oct 6, 1998·Annual Review of Biochemistry·S Verma, F Eckstein
Apr 1, 1997·Proceedings of the National Academy of Sciences of the United States of America·S A Strobel, K Shetty
Dec 14, 2011·Chembiochem : a European Journal of Chemical Biology·Lasse H LauridsenRakesh N Veedu
Jun 11, 1999·Current Opinion in Structural Biology·S A Strobel
Apr 20, 1999·Methods : a Companion to Methods in Enzymology·S P Ryder, S A Strobel
May 21, 2016·Frontiers in Chemistry·María A DellafioreAdolfo M Iribarren
Sep 6, 2000·Journal of Molecular Biology·J K Strauss-Soukup, S A Strobel
Oct 4, 2012·Nucleic Acid Therapeutics·Partha RayRebekah R White
Apr 23, 2008·Current Protocols in Nucleic Acid Chemistry·Jesse C Cochrane, Scott A Strobel
May 12, 2009·Acta Biochimica Et Biophysica Sinica·Zhifeng ZengTianhong Zhou
Jan 29, 2019·Artificial Cells, Nanomedicine, and Biotechnology·Bhagwan S BatuleHyun Gyu Park

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