Enzymatic RNA synthesis with deoxynucleoside 5'-O-(1-thiotriphosphates).
We have investigated the incorporation of 2'-deoxynucleoside-5'-O-(1-thiotriphosphates) into RNA transcripts using T7 RNA polymerase. With the exception of [alpha-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, [alpha-S]dTTP and [alpha-S]dATP, were inserted specifically. However, [alpha-S]dCTP was excluded at specific sites. We could not obtain transcripts using the deoxyguanosine derivative.
Gaur
R K
RK
Institut für Allgemeine Mikrobiologie, Christian-Albrechts-Universität Kiel, Germany.
Krupp
G
G
D001483:Base Sequence; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism*; D003854:Deoxyribonucleotides / Q000378:metabolism*; D006860:Hydrogen Bonding; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D009694:Nucleic Acid Precursors / Q000737:chemistry; D012313:RNA / Q000737:chemistry*; D012323:RNA Processing, Post-Transcriptional; D012360:RNA, Transfer, Phe / Q000737:chemistry; D013873:Thionucleotides / Q000378:metabolism*; D014764:Viral Proteins
D003854:Deoxyribonucleotides; D009694:Nucleic Acid Precursors; D012360:RNA, Transfer, Phe; D013873:Thionucleotides; D014764:Viral Proteins; D012313:RNA; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases