Enzymatic RNA synthesis with deoxynucleoside 5'‐O‐(1‐thiotriphosphates)

FEBS Letters

Krupp, Guido¹  Gaur, Rajesh K.¹ 

[1] Institut für Allgemeine Mikrobiologie, Christian-Albrechts-Universität Kiel, Am Botanischen Garten 9, W-2300 Kiel 1, Germany

关键词: T7 RNA polymerase;    Transcription;    in vitro;    Modification interference;    Pre-tRNA;    Modified nucleotide;    [α-S]NTP;    ribonucleoside 5'-O-(1-thiotriphosphate);    [α-S]dNTP;    2'-deoxynucleoside-5'-O-(1-thiotriphosphate);    NTPs;    nucleoside triphosphates;    dNTPs;    deoxynucleoside triphosphates.;   

DOI  :  10.1016/0014-5793(93)81132-J

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

We have investigated the incorporation of 2'-deoxynucleoside-5'-O-(1-thiotriphosphates) into RNA transcripts using T7 RNA polymerase. With the exception of [α-S]dGTP, we obtained full-length transcripts of pre-tRNA^phe and pre-tRNA^tyr using an appropriate mixture of 2'-deoxynucleoside 5'-O-(1-thiotriphosphate) and the corresponding normal nucleoside triphosphate. The yields of the transcripts were comparable to those obtained with unmodified NTPs. Both substrates, [α-S]dTTP and [α-S]dATP, were inserted specifically. However, [α-S]dCTP was excluded at specific sites. We could not obtain transcripts using the deoxyguanosine derivative.

【 授权许可】

Unknown