Reverse transcription of tRNA.
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)-tRNA3Gly(E.coli). The transcription products were characterized by sequence analysis.
D000227:Adenine Nucleotides; D001483:Base Sequence; D004247:DNA / Q000096:biosynthesis*; D004926:Escherichia coli; D006157:Guanosine Monophosphate / Q000378:metabolism; D009691:Nucleic Acid Denaturation; D009841:Oligonucleotides / Q000378:metabolism; D012343:RNA, Transfer / Q000378:metabolism*; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism; D012441:Saccharomyces cerevisiae
D000227:Adenine Nucleotides; D009841:Oligonucleotides; D006157:Guanosine Monophosphate; D004247:DNA; D012343:RNA, Transfer; D012194:RNA-Directed DNA Polymerase