Mobilization of quiet, endogenous Tc3 transposons of Caenorhabditis elegans by forced expression of Tc3 transposase.
The commonly studied Caenorhabditis elegans strain Bristol N2 contains approximately 15 copies per genome of the transposon Tc3. However, Tc3 is not active in Bristol N2. Tc3 contains one major open reading frame (Tc3A). We have fused this open reading frame to an inducible promoter and expressed it in a transgenic Bristol N2 line. Tc3A expression resulted in frequent excision and transposition of endogenous Tc3 elements. This shows that the Bristol N2 genome contains Tc3 transposons that are cis proficient for transposition, but are immobile because Tc3A is absent. We demonstrate that recombinant Tc3A binds specifically to the terminal nucleotides of the Tc3 inverted repeat, indicating that Tc3A is the Tc3 transposase. Activation of Tc3 transposition in vivo was accompanied by the appearance of extrachromosomal, linear copies of Tc3. These may be intermediates in Tc3 transposition.
Netherlands Cancer Institute, Division of Molecular Biology, Amsterdam.
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van Luenen
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Colloms
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Plasterk
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D017173:Caenorhabditis elegans / Q000201:enzymology / Q000235:genetics*; D002875:Chromosomes; D003001:Cloning, Molecular; D004247:DNA / Q000378:metabolism; D004251:DNA Transposable Elements; D015870:Gene Expression; D007814:Larva / Q000254:growth & development; D008969:Molecular Sequence Data; D009713:Nucleotidyltransferases / Q000378:metabolism*; D012091:Repetitive Sequences, Nucleic Acid; D019895:Transposases
D004251:DNA Transposable Elements; D004247:DNA; D009713:Nucleotidyltransferases; D019895:Transposases