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Nucleic Acids Research, 1978, Vol. 5, No. 10 3821-3830
© 1978

Articles

Preparation of synthetic tRNA precursors with tRNA nuc1eotidy1transferase*

Murray P. Deutscher and Ranajit K. Ghosh+

Department of Biochemistry, University of Connecticut Health Center Farmington, CT 06032, USA +Indian Institute of Experimental Medicine Jadavpur, Calcutta, India

Received July 27, 1978.

Rabbit liver tRNA nucleotidyltransferase can be used to substitute nucleotides within the -C-C-A sequence of tRNA or to add nucleotides following this sequence. These anomolous reactions of the enzyme have been used to prepare radioactively-labeled synthetic tRNA precursors which mimic the structure of the natural precursors. Under appropriate conditions synthetic precursors of defined structure can be made. In this paper we describe the synthesis of tRNA-C-[14C]U and tRNA-C-C-A-[14C]-C, which are representative of tRNA precursors containing altered residues within the -C-C-A sequence or with extra residues following the normal 3'terminus. A variety of other possible precursors can also be prepared. These synthetic tRNA precursors have already proved useful for isolation of possible tRNA processing nucleases.

*This paper is number 24 in the series, "Reactions at the 3'Terminus of tRNA". The previous paper in the series is reference 23.


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