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. 1980 Sep 11;8(17):3909–3916. doi: 10.1093/nar/8.17.3909

Comparison of substrate base sequences for RNA ligase reactions in the synthesis of a tetradecanucleotide corresponding to bases 21-34 of E. coli tRNAfMet 1.

E Ohtsuka, T Doi, H Uemura, Y Taniyama, M Ikehara
PMCID: PMC324203  PMID: 7003540

Abstract

A tetradecanucleotide U-A-G-C(U-C-G)2G-G-C-Up corresponding to bases 21-34 of a nascent sequence of formylmethionyl tRNA of E. coli has been synthesized by the joining of two combinations of chemically synthesized oligonucleotides: 1) U-A-G-C + U-C-G-U-C-G + G-G-C-Up and 2) U-A-G-C + U-C-G-U + C-G-G-G-C-Up. In reaction 1) and the extent of joining *pG-G-C-Up to U-C-G-U-C-G was only 15.4% and the last ligation of the decamer to U-A-G-U proceeded to 27%. In reaction 2) joining between U-A-G-C and pU-C-G-Up gave a high yield (88%). The ligation of this octamer and *pC-G-G-G-C-Up also gave a satisfactory yield (52%). These reactions suggest that sequence preferences in RNA ligase reactions may arise from the structure of the 3'-end of acceptor molecules.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Brahms J., Aubertin A. M., Dirheimer G., Grunberg-Manago M. Studies of trinucleotide conformations. Role of guanine residues in an oligonucleotide chain. Biochemistry. 1969 Aug;8(8):3269–3278. doi: 10.1021/bi00836a021. [DOI] [PubMed] [Google Scholar]
  2. Brownlee G. G., Sanger F. Chromatography of 32P-labelled oligonucleotides on thin layers of DEAE-cellulose. Eur J Biochem. 1969 Dec;11(2):395–399. doi: 10.1111/j.1432-1033.1969.tb00786.x. [DOI] [PubMed] [Google Scholar]
  3. Jay E., Bambara R., Padmanabhan R., Wu R. DNA sequence analysis: a general, simple and rapid method for sequencing large oligodeoxyribonucleotide fragments by mapping. Nucleic Acids Res. 1974 Mar;1(3):331–353. doi: 10.1093/nar/1.3.331. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Ohtsuka E., Nishikawa S., Fukumoto R., Tanaka S., Markham A. F. Joining of synthetic ribotrinucleotides with defined sequences catalyzed by T4 RNA ligase. Eur J Biochem. 1977 Dec 1;81(2):285–291. doi: 10.1111/j.1432-1033.1977.tb11950.x. [DOI] [PubMed] [Google Scholar]
  5. Ohtsuka E., Nishikawa S., Fukumoto R., Uemura H., Tanaka T., Nakagawa E., Miyake T., Ikehara M. Synthesis of 5' fragments of formylmethionine transfer ribonucleic acid and their reconstitution with a natural three-quarter molecule. Eur J Biochem. 1980 Apr;105(3):481–487. doi: 10.1111/j.1432-1033.1980.tb04523.x. [DOI] [PubMed] [Google Scholar]
  6. Ohtsuka E., Nishikawa S., Markham A. F., Tanaka S., Miyake T., Wakabayashi T., Ikehara M., Sugiura M. Joining of 3'-modified oligonucleotides by T4 RNA ligase. Synthesis of a heptadecanucleotide corresponding to the bases 61--77 from Escherichia coli tRNAfMet. Biochemistry. 1978 Nov 14;17(23):4894–4899. doi: 10.1021/bi00616a006. [DOI] [PubMed] [Google Scholar]
  7. Ohtsuka E., Nishikawa S., Sugiura M., Ikehara M. Joining of ribooligonucleotides with T4 RNA ligase and identification of the oligonucleotide-adenylate intermediate. Nucleic Acids Res. 1976 Jun;3(6):1613–1623. doi: 10.1093/nar/3.6.1613. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Sugiura M., Suzuki M., Ohtsuka E., Nishikawa S., Uemura H., Ikehara M. Purification of T4 RNA ligase by 2', 5'-ADP sepharose chromatography. FEBS Lett. 1979 Jan 1;97(1):73–76. doi: 10.1016/0014-5793(79)80055-1. [DOI] [PubMed] [Google Scholar]

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