Cis requirements for transposition of Tc1-like transposons in C. elegans.
The Caenorhabditis elegans transposons Tc1 and Tc3 are able to transpose in heterologous systems such as human cell lines and zebrafish. Because these transposons might be useful vectors for transgenesis and mutagenesis of diverse species, we determined the minimal cis requirements for transposition. Deletion mapping of the transposon ends shows that fewer than 100 bp are sufficient for transposition of Tc3. Unlike Tc1, Tc3 has a second, internal transposase binding site at each transposon end. We found that these binding sites play no major role in the transposition reaction, since they can be deleted without reduction of the transposition frequency. Site-directed mutagenesis was performed on the conserved terminal base pairs at the Tc3 ends. The four terminal base pairs at the ends of the Tc3 inverted repeats were shown to be required for efficient transposition. Finally, increasing the length of the transposon from 1.9 kb to 12.5 kb reduced the transposition frequency by 20-fold, both in vivo and in vitro.
Fischer
S E
SE
Division of Molecular Biology and Center for Biomedical Genetics, The Netherlands Cancer Institute, Amsterdam.
van Luenen
H G
HG
Plasterk
R H
RH
D000818:Animals; D030801:Animals, Genetically Modified; D001665:Binding Sites; D017173:Caenorhabditis elegans / Q000201:enzymology* / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics*; D017238:Genes, Helminth; D016296:Mutagenesis; D012091:Repetitive Sequences, Nucleic Acid; D019895:Transposases / Q000235:genetics*
D004268:DNA-Binding Proteins; C081454:Tc1 transposase; D019895:Transposases