RRC ID 4802
Author Shibano T, Takeda M, Suetake I, Kawakami K, Asashima M, Tajima S, Taira M.
Title Recombinant Tol2 transposase with activity in Xenopus embryos.
Journal FEBS Lett.
Abstract The Tol2 transposon system is a useful gene transduction technique, but the injection of mRNA is not sufficiently effective in Xenopus embryos to express Tol2 transposase (Tol2TP). To overcome this, we bacterially synthesized recombinant Tol2TP (rTol2TP) protein and showed that rTol2TP efficiently excised the Tol2 element from an injected donor plasmid in Xenopus embryos. Furthermore, injected embryos exhibited uniform and ubiquitous expression of an EGFP reporter gene placed within the Tol2 element. Importantly, size-exclusion chromatography suggests that rTol2TP forms a tetramer, which differs from the reported hexamer formed by Hermes transposase, although both belong to the same hAT family. The use of rTol2TP may facilitate efficient gene transduction in Xenopus, and the biochemical characterization of Tol2TP.
Volume 581(22)
Pages 4333-6
Published 2007-9-4
DOI 10.1016/j.febslet.2007.08.004
PII S0014-5793(07)00871-X
PMID 17716667
MeSH Animals Base Sequence Chromatography, Gel DNA Transposable Elements / genetics Escherichia coli / genetics Green Fluorescent Proteins / genetics Green Fluorescent Proteins / metabolism Microinjections Oryzias / genetics Recombinant Proteins / biosynthesis Recombinant Proteins / genetics Recombinant Proteins / isolation & purification Recombinant Proteins / metabolism* Transformation, Genetic Transposases / biosynthesis Transposases / genetics Transposases / isolation & purification Transposases / metabolism* Xenopus / classification Xenopus / embryology* Xenopus / genetics Xenopus / metabolism*
IF 2.999
Times Cited 15
Xenopus tropicalis