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...

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Bibliographic Details
Published in:Genes & development 1996-03, Vol.10 (6), p.755-761
Main Authors: Vos, J C, De Baere, I, Plasterk, R H
Format: Article
Language:English
Subjects:
Animals
Animals, Genetically Modified
Base Sequence
Blotting, Southern
Caenorhabditis elegans
Caenorhabditis elegans - enzymology
Caenorhabditis elegans - genetics
Cell Nucleus - metabolism
Cell-Free System
DNA Primers
DNA Transposable Elements
DNA-Binding Proteins - genetics
DNA-Binding Proteins - metabolism
Drug Resistance - genetics
Escherichia coli - enzymology
Escherichia coli - genetics
Models, Genetic
Molecular Sequence Data
Mutation
Nucleotidyltransferases - genetics
Nucleotidyltransferases - metabolism
Polymerase Chain Reaction
Recombinant Proteins - isolation & purification
Sequence Analysis, DNA
Transposases
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Summary: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.
ISSN:0890-9369
1549-5477
DOI:10.1101/gad.10.6.755