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Enzymatic RNA synthesis with deoxynucleoside 5'‐ O ‐(1‐thiotriphosphates)
Author(s) -
Gaur Rajesh K.,
Krupp Guido
Publication year - 1993
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(93)81132-j
Subject(s) - rna , nucleoside , deoxyguanosine , transfer rna , polymerase , chemistry , enzyme , biochemistry , rna polymerase , microbiology and biotechnology , t7 rna polymerase , nucleotide , dna , stereochemistry , biology , gene , escherichia coli , bacteriophage
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.