Biopolymers and cell. 1990. Volume 6. № 3. 50 - 55

L. Ya. Denisova, S. N. Zagrebelny, N. M. Pustoshilova, A. G. Venyaminova, V. V. Corn, V. F. Zaritova, M. N. Repkova

PREPARATIVE SYNTHESIS OF RIBOOLIGONUCLEOTIDES WITH THE PRESET SEQUENCE BY TRANSCRIPTION OF DEOXYRIBONUCLEOTIDES

Summary

    The transcription of complementary deoxyribooligonucleotides was performed to obtain AGGGGAUUGAAAAUC (tRNAphe anticodon loop fragment) and AUGAGGAAUACCCAUG (the MS2 phase RNA fragment) in preparative amounts. The nucleotide incorporation into the transcription product was shown to achieve 70 % of the template nucleotide level. The procedure was developed for the isolation of the transcript from the reaction mixture. The isolation steps involve geparine-agarose chromatography, Lichroprep RP 18 one and the electrophoresis in 20 % polyacrylamide gel under denaturation conditions followed by electroelution of the desired product from gel. Being an exact copy of deoxyribotemplate the ribooligonucleotide gave the yield of 1.0-1.5 U26o on 5.0 U2eo of the template.