Transposase is the only nematode protein required for in vitro transposition of Tc1.
The Tc1 element of Caenorhabditis elegans is a member of the most widespread class of DNA transposons known in nature. Here, we describe efficient and precise transposition of Tc1 in a cell-free system. Tc1 appears to jump by a cut-and-paste mechanism of transposition. The terminal 26 bp of the Tc1 terminal repeats together with the flanking TA sequence are sufficient for transposition. The target site choice in vitro is similar to that in vivo. Transposition is achieved with an extract prepared from nuclei of transgenic nematodes that overexpress Tc1 transposase but also by recombinant transposase purified from Escherichia coli. The simple reaction requirements explain why horizontal spread of Tc1/mariner transposons can occur. They also suggest that Tcl may be a good vector for transgenesis of diverse animal species.
Vos
J C
JC
The Netherlands Cancer Institute, Division of Molecular Biology, Amsterdam.
De Baere
I
I
Plasterk
R H
RH
D000818:Animals; D030801:Animals, Genetically Modified; D001483:Base Sequence; D015139:Blotting, Southern; D017173:Caenorhabditis elegans / Q000201:enzymology / Q000235:genetics*; D002467:Cell Nucleus / Q000378:metabolism; D002474:Cell-Free System; D017931:DNA Primers; D004251:DNA Transposable Elements; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D004351:Drug Resistance / Q000235:genetics; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D008957:Models, Genetic; D008969:Molecular Sequence Data; D009154:Mutation; D009713:Nucleotidyltransferases / Q000235:genetics / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000302:isolation & purification; D017422:Sequence Analysis, DNA; D019895:Transposases
D017931:DNA Primers; D004251:DNA Transposable Elements; D004268:DNA-Binding Proteins; D011994:Recombinant Proteins; D009713:Nucleotidyltransferases; C081454:Tc1 transposase; D019895:Transposases