title: Establishment of transgenic lines for jumpstarter method using a composite transposon vector in the ladybird beetle, Harmonia axyridis.
abstract: In this post-genomic era, genome-wide functional analysis is indispensable. The recent development of RNA interference techniques has enabled researchers to easily analyze gene function even in non-model organisms. On the other hand, little progress has been made in the identification and functional analyses of cis-regulatory elements in non-model organisms. In order to develop experimental platform for identification and analyses of cis-regulatory elements in a non-model organism, in this case, the ladybird beetle, Harmonia axyridis, we established transgenic transposon-tagged lines using a novel composite vector. This vector enables the generation of two types of insertion products (jumpstarter and mutator). The jumpstarter portion carries a transposase gene, while the mutator segment carries a reporter gene for detecting enhancers. The full-composite element is flanked by functional termini (required for movement); however, the mutator region has an extra terminus making it possible for the mutator to remobilize on its own, thus leaving an immobile jumpstarter element behind. Each insertion type is stable on its own, but once crossed, jumpstarters can remobilize mutators. After crossing a jumpstarter and mutator line, all tested G2 females gave rise to at least one new insertion line in the next generation. This jumping rate is equivalent to the P-element-mediated jumpstarter method in Drosophila. These established transgenic lines will offer us the ideal experimental materials for the effective screening and identification of enhancers in this species. In addition, this jumpstarter method has the potential to be as effective in other non-model insect species and thus applicable to any organism.
authors: Graduate School of Bioagricultural Sciences, Nagoya University, Chikusa, Nagoya, Japan.
authors: Hisashi
authors: H
authors: Kuwayama
authors: Graduate School of Bioagricultural Sciences, Nagoya University, Chikusa, Nagoya, Japan; Department of Entomology, Washington State University, Pullman, Washington, United States of America.
authors: Hiroki
authors: H
authors: Gotoh
authors: Graduate School of Bioagricultural Sciences, Nagoya University, Chikusa, Nagoya, Japan.
authors: Yusuke
authors: Y
authors: Konishi
authors: Graduate School of Bioagricultural Sciences, Nagoya University, Chikusa, Nagoya, Japan.
authors: Hideto
authors: H
authors: Nishikawa
authors: Graduate School of Bioagricultural Sciences, Nagoya University, Chikusa, Nagoya, Japan.
authors: Toshinobu
authors: T
authors: Yaginuma
authors: Graduate School of Bioagricultural Sciences, Nagoya University, Chikusa, Nagoya, Japan.
authors: Teruyuki
authors: T
authors: Niimi
mesh_terms: D000818:Animals; D001517:Coleoptera / Q000235:genetics*; D004251:DNA Transposable Elements; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors; D016297:Mutagenesis, Site-Directed / Q000379:methods; D030781:Organisms, Genetically Modified; D019895:Transposases / Q000235:genetics
chemical_list: D004251:DNA Transposable Elements; D019895:Transposases