RRC ID 207
Author Kawakami K, Imanaka K, Itoh M, Taira M.
Title Excision of the Tol2 transposable element of the medaka fish Oryzias latipes in Xenopus laevis and Xenopus tropicalis.
Journal Gene
Abstract The Tol2 transposable element from the medaka fish belong to the hAT family of transposons. In the previous studies, we have identified an autonomous member of this element, which encodes a fully functional transposase, and have shown that it can catalyze transposition in the zebrafish germ lineage. To date, the Tol2 element is the only natural transposon in vertebrates from which an autonomous member has been identified. We report here transposase-dependent excision of the Tol2 element in Xenopus laevis and Xenopus (Silurana) tropicalis embryos. We coinjected a plasmid DNA containing a nonautonomous Tol2 element and the transposase mRNA synthesized in vitro into two-cell-stage embryos, and analyzed DNA extracted from the injected embryos by polymerase chain reaction (PCR). We demonstrated that the Tol2 element could be excised from the plasmid DNA in both X. laevis and X. tropicalis only when it was coinjected with the transposase mRNA. In most cases, a complete loss of the Tol2 sequence was accompanied by addition of a short DNA sequence to the target sequence, indicating that transposase-dependent excision occurred. While these footprints were characteristic to those created upon excision of transposons of the hAT family, the additional bases found in Xenopus were longer and their structures were more complicated than those detected upon excision in zebrafish. This may reflect differences in the activities of host factors involved in either transposition, repair, or both between fish and frog. Our present study suggests that the Tol2 transposon system should be used as a novel genetic tool to develop transgenesis and mutagenesis methods in Xenopus.
Volume 338(1)
Pages 93-8
Published 2004-8-18
DOI 10.1016/j.gene.2004.05.013
PII S037811190400304X
PMID 15302410
MeSH Animals Base Sequence DNA / chemistry DNA / genetics DNA Transposable Elements / genetics* Embryo, Nonmammalian / metabolism* Female Microinjections Mutagenesis, Insertional Oryzias / genetics Plasmids / administration & dosage Plasmids / genetics Polymerase Chain Reaction RNA, Messenger / administration & dosage RNA, Messenger / genetics Sequence Analysis, DNA Transposases / genetics Xenopus Xenopus laevis
IF 2.984
Times Cited 39
Clawed frogs / Newts