Post-integration stabilization of a transposon vector by terminal sequence deletion in Drosophila melanogaster

被引:50
作者
Handler, AM
Zimowska, GJ
Horn, C
机构
[1] USDA ARS, Ctr Med Agr & Vet Entomol, Gainesville, FL 32608 USA
[2] Max Planck Inst Mol Genet, Cardiovasc Res Ctr, D-10115 Berlin, Germany
关键词
D O I
10.1038/nbt1002
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Germline transformation systems for nearly 20 insect species have been derived from transposable elements, allowing the development of transgenic insects for basic and applied studies(1-3). These systems use a defective nonautonomous vector that results in stable vector integrations after the disappearance of transiently provided transposase helper plasmid(4), which is essential to maintain true breeding lines and consistent transgene expression that would otherwise be lost after vector remobilization. The risk of remobilization by an unintended transposase source has so far not been a concern for laboratory studies, but the prospective use of millions of transgenic insects in biocontrol programs will likely increase the risk, therefore making this a critical issue for the ecological safety of field release programs(5,6). Here we describe an efficient method that deletes a terminal repeat sequence of a transposon vector after genomic integration. This procedure prevents transposase-mediated remobilization of the other terminal sequence and associated genes, ensuring their genomic stability.
引用
收藏
页码:1150 / 1154
页数:5
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