RESUMEN

The lepidopteran transposable element piggyBac can mediate germline insertions in at least four insect orders. It therefore shows promise as a broad-spectrum transformation vector, but applications such as enhancer trapping and transposon-tag mutagenesis are still lacking. We created, cloned, sequenced and genetically mapped a set of piggyBac insertions in the red flour beetle, Tribolium castaneum. Transpositions were precise, and specifically targeted the canonical TTAA recognition sequence. We detected several novel reporter-expression domains, indicating that piggyBac could be used to identify enhancer regions. We also demonstrated that a primary insertion of a non-autonomous element can be efficiently remobilized to non-homologous chromosomes by injection of an immobile helper element into embryos harbouring the primary insertion. These developments suggest potential for more sophisticated methods of piggyBac-mediated genome manipulation.

Asunto(s)

Elementos Transponibles de ADN/genética , Transformación Genética , Tribolium/genética , Animales , Secuencia de Bases , Southern Blotting , Mapeo Cromosómico , Perfilación de la Expresión Génica , Microinyecciones , Datos de Secuencia Molecular , Plásmidos

Asunto(s)

Marcadores Genéticos , Proteínas de Homeodominio , Insectos/genética , Animales , Animales Modificados Genéticamente , Elementos Transponibles de ADN , Proteínas de Unión al ADN/genética , Drosophila/genética , Proteínas del Ojo , Técnicas de Transferencia de Gen , Proteínas Fluorescentes Verdes , Proteínas Luminiscentes/genética , Factor de Transcripción PAX6 , Factores de Transcripción Paired Box , Regiones Promotoras Genéticas , Proteínas Represoras , TATA Box , Tribolium/genética