Sperm-mediated transformation of the honey bee, Apis mellifera

Our primary objective was to identify techniques to transform the genome of the honey bee (Apis mellifera) with foreign DNA constructs. The strategy we adopted wes to linearize foreign DNA and introduce it with sperm during the instrumental insemination of virgin queen honey bees. We analysed extracts from larvae within the same cohort end isolated the predicted fragment by means of PCR amplification of genomic DNA. Larvae that carried the construct also expressed the introduced DNA. We propagated several trensgenic lines for up to three generations, which demonstrates its heritability. Once carried by e queen, the construct can be detected in that queen's larvae over several months. However, there wes no evidence of integration of the construct, et least es determined by genomic Southern analysis. Nevertheless, this demonstrates the general viability of the technique for introduction of DNA, end it should be augmented by further use of transposable elements that enhance integration.