A versatile vector set for animal transgenesis

Genetic manipulation of a series of diverged arthropods is a highly desirab le goal for a better understanding of developmental and evolutionary proces ses. A major obstacle so far has been the difficulty in obtaining marker ge nes that allow easy and reliable identification of transgenic animals. Here , we present a versatile vector set for germline transformation based on th e promiscuous transposons mariner, Hermes and piggyBac. Into these vectors, we introduced a potentially universal marker system that is comprised of a n artificial promoter containing three Pax-6 homodimer binding sites. This promoter drives strong expression of spectral variants of the enhanced gree n fluorescent protein (EGFP) in larval, pupal, and adult photoreceptors. Us ing special filter sets, the yellow (EYFP) and cyan (ECFP) variant are full y distinguishable and therefore represent a separable pair of markers. Furt hermore, we adapted a simple plasmid-based transposition assay system to en able quick functional tests of our vectors in different arthropod species b efore employing them in more laborious germline transformation experiments. Using this system we demonstrate that our vectors transpose in both Drosop hila melanogaster and Drosophila virilis.