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Nucleic Acids Research, 1978, Vol. 5, No. 4 1165-1178
© 1978

Articles

Reverse transcription of tRNA

Burghardt Wittig and Stephanie Wittig

Institut für Molekularbiologie und Biochemie, Freie Universität Berlin Arnimallee 22, D-1000 Berlin 33, GFR

Received February 13, 1978. The 3' terminus of tRNA was enzymatically elongated by an oligo(A) tail. A fragment of DNA polymerase I (E.coli) was used in the presence of manganese to phase and synthesize a cleavable primer at the oligo(A)-tRNA template. When the threedimensional structure of oligo(A)-tRNA is being unfolded under conditions where the primer is still hybridized at the oligo(A) tail, the DNA polymerase I fragment transcribes oligo(A)-tRNA into DNA. Reverse transcription is slowed down and its fidelity suspended by the 1-methyladenine in oligo(A)-tRNAPhe(yeast). The reaction is stopped by the highly modified Y-base present in this template. Approximately full length transcripts can be obtained from oligo(A)-Formula(E.coli). The transcription products were characterized by sequence analysis.


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