Abstract
Germ line transformation is a powerful research tool to probe and manipulate gene function but currently the number of insect species transformed is small and biased towards Diptera. Here we develop transgenic techniques for a butterfly in the family Pieridae, the widely-studied cabbage white butterfly (Pieris rapae L; Lepidoptera: Pieridae). Germ line transformation of pierids is useful because butterfly wing patterns have become model systems in evolutionary developmental biology, and the previously transformed butterflies are members of a different family, the nymphalids. We used a piggyBac[3xP3-EGFP] construct as a marker for germ line transformation and obtained a single transformed adult that did not reproduce but which displayed fluorescence in its eyes. After identifying the genomic insertion site for the construct we conclude that this individual was likely transgenic. However, short of longer-term inheritance data, we discuss two less likely alternative possibilities for our data.
Highlights
Wing patterns are becoming important model systems for addressing questions in evolutionary developmental biology [1,2,3], to examine the evolution of gene function or regulation in butterflies it will be important to perform germ-line transformation in several of these insects.Two species of butterfly, Bicyclus anynana and Danaus plexippus have been genetically transformed using piggyBac transposable elements and zinc-finger nucleases [4,5], respectively
We used a piggyBac construct containing the [3xP3-EGFP] marker cassette that upon insertion into the germ line led to green fluorescence in the eyes of Pieris rapae
We used TAIL-PCR [11] to isolate the left and right genomic sequences flanking the piggyBac insertion in individuals that were positive for EGFP
Summary
Wing patterns are becoming important model systems for addressing questions in evolutionary developmental biology (evo-devo) [1,2,3], to examine the evolution of gene function or regulation in butterflies it will be important to perform germ-line transformation in several of these insects.Two species of butterfly, Bicyclus anynana and Danaus plexippus have been genetically transformed using piggyBac transposable elements and zinc-finger nucleases [4,5], respectively. We used a piggyBac construct containing the [3xP3-EGFP] marker cassette that upon insertion into the germ line led to green fluorescence in the eyes of Pieris rapae.
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