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Nucleic Acids Research, 1980, Vol. 8, No. 3 601-610
© 1980

Articles

Elongation of oligonucleotides in the 3'-direction with activated mononucleotides and their analogs using RNA ligase

E. Ohtsuka, T. Miyake, K. Nagao, H. Uemura, S. Nishikawa, K. Nagao, M. Sugiura and M. Ikehara

Department of Biochemistry, Cancer Institute, Japanese Foundation for Cancer Research Kami-Ikebukuro, Toshima-ku, Tokyo, 170, Japan

Received December 5, 1979. P1-Adenosine 5'-P2 ,3'-ethoxymethylidene nucleosides [A(5')ppN from four common nucleosides have been prepared and used for single addition of nucleotides to elongate oligo-nucleotide chains in the 3'-direction in RNA ligase reaction. U-U-C, T-Y-C and A-C-C were used as acceptors. Structural dependence in these acceptors was found to be smaller compared to joining reactions between oligonucleotides. Adenosine analogs including 8-bromo-, 2'-fluoro-, 2'-azido-, 8,2'-O-cyclo-, 8,2'- S-cyclo-adenosine, arabinosyladenine and 2'-deoxyadenosine were added to the 3'-end of A-C-C by adenylation chemically followed by joining with RNA ligase. Symmetrical 5'-pyrophosphates of 8-bromo-, 2'-fluoro- and 2'-azido-adenosine were not recognized as donor substrates.


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