Demonstration of retrotransposition of the Tf1 element in fission yeast.
Tf1, a retrotransposon from fission yeast, has LTRs and coding sequences resembling the protease, reverse transcriptase and integrase domains of retroviral pol genes. A unique aspect of Tf1 is that it contains a single open reading frame whereas other retroviruses and retrotransposons usually possess two or more open reading frames. To determine whether Tf1 can transpose, we overproduced Tf1 transcripts encoded by a plasmid copy of the element marked with a neo gene. Approximately 0.1-4.0% of the cell population acquired chromosomally inherited resistance to G418. DNA blot analysis demonstrated that such strains had acquired both Tf1 and neo specific sequences within a restriction fragment of the same size; the size of this restriction fragment varied between different isolates. Structural analysis of the cloned DNA flanking the Tf1-neo element of two transposition candidates with the same regions in the parent strain showed that the ability to grow on G418 was due to transposition of Tf1-neo and not other types of recombination events.
Levin
H L
HL
Department of Molecular Biology and Genetics, Johns Hopkins School of Medicine, Baltimore, MD 21205.
Boeke
J D
JD
D001483:Base Sequence; D015139:Blotting, Southern; D015342:DNA Probes; D004251:DNA Transposable Elements; D004271:DNA, Fungal / Q000235:genetics; D005800:Genes, Fungal; D008969:Molecular Sequence Data; D010957:Plasmids; D011401:Promoter Regions, Genetic; D012331:RNA, Fungal / Q000235:genetics; D012091:Repetitive Sequences, Nucleic Acid; D012441:Saccharomyces cerevisiae / Q000235:genetics; D012568:Schizosaccharomyces / Q000235:genetics*; D013045:Species Specificity; D013696:Temperature; D014158:Transcription, Genetic
D015342:DNA Probes; D004251:DNA Transposable Elements; D004271:DNA, Fungal; D012331:RNA, Fungal