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

AbstractWe 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-tRNAphe and pre-tRNAtyr 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