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Synthesis of deoxynucleotide-containing RNA-like single-stranded polynucleotides (dcRNAs) using the Y639F, S641A mutant of T7 RNA polymerase (T7 RNAP) was studied. A number of different T7 promoter-containing plasmids were tested as templates for dcRNA synthesis. The dcRNA synthesis efficiency strongly depended on the sequence of the first 8-10 nucleotides immediately downstream of the promoter and increased with the distance of the first incorporated dNMP from the transcription start. The incorporation of dGMP which is obligatory for most T7 promoters in positions +1-+2(3) was practically negligible. Using the constructed plasmid pTZR7G containing seven dG links in the non-coding chain immediately downstream of the promoter, the synthesis of all possible dcRNAs (except dG-containing) was achieved with high yields.
S O Gudima, and
D A Kostyuk, and
O I Grishchenko, and
V L Tunitskaya, and
L V Memelova, and
S N Kochetkov
July 1994,
Proceedings of the National Academy of Sciences of the United States of America,
S O Gudima, and
D A Kostyuk, and
O I Grishchenko, and
V L Tunitskaya, and
L V Memelova, and
S N Kochetkov
January 2004,
Molekuliarnaia biologiia,
S O Gudima, and
D A Kostyuk, and
O I Grishchenko, and
V L Tunitskaya, and
L V Memelova, and
S N Kochetkov
September 1995,
The EMBO journal,
S O Gudima, and
D A Kostyuk, and
O I Grishchenko, and
V L Tunitskaya, and
L V Memelova, and
S N Kochetkov
April 1986,
Proceedings of the National Academy of Sciences of the United States of America,
S O Gudima, and
D A Kostyuk, and
O I Grishchenko, and
V L Tunitskaya, and
L V Memelova, and
S N Kochetkov
July 1973,
Biochimica et biophysica acta,
S O Gudima, and
D A Kostyuk, and
O I Grishchenko, and
V L Tunitskaya, and
L V Memelova, and
S N Kochetkov
April 1995,
Doklady Akademii nauk,
S O Gudima, and
D A Kostyuk, and
O I Grishchenko, and
V L Tunitskaya, and
L V Memelova, and
S N Kochetkov
March 2001,
Chembiochem : a European journal of chemical biology,
S O Gudima, and
D A Kostyuk, and
O I Grishchenko, and
V L Tunitskaya, and
L V Memelova, and
S N Kochetkov
October 1993,
European journal of biochemistry,
S O Gudima, and
D A Kostyuk, and
O I Grishchenko, and
V L Tunitskaya, and
L V Memelova, and
S N Kochetkov
April 1987,
Cell,
S O Gudima, and
D A Kostyuk, and
O I Grishchenko, and
V L Tunitskaya, and
L V Memelova, and
S N Kochetkov
August 1980,
The Journal of biological chemistry,
