Germline transformation of Drosophila melanogaster with the piggyBac transposon vector

Authors

  • A. M. Handler,

    Corresponding author
    1. Center for Medical, Agricultural, and Veterinary Entomology, Agricultural Research Service, US Department of Agriculture, Gainesville, FL, U.S.A.
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  • R. A. Harrell Ii

    1. Center for Medical, Agricultural, and Veterinary Entomology, Agricultural Research Service, US Department of Agriculture, Gainesville, FL, U.S.A.
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Dr Alfred Handler, USDA-ARS, 1700 SW 23rd Drive, Gainesville, FL 32608, U.S.A. E-mail: handler@nersp.nerdc.ufl.edu

Abstract

Germline transformation of Drosophila melanogaster was attempted with the piggyBac gene-transfer system from the cabbage looper moth, Trichoplusia ni. Using a self-regulated transposase helper and a white marked vector, a transformation frequency of 1–3% per fertile G0 was obtained, similar to that previously achieved in the medfly. Use of an hsp70-regulated helper increased this frequency more than eight-fold. Transformation with a vector marked with white and green fluorescent protein (GFP) under polyubiquitin–nuclear localizing sequence regulation yielded seventy G1 transformants which all expressed GFP, but only twenty-seven of these expressed eye pigmentation that would have allowed their selection based on white+ expression. PiggyBac transformation in two distantly related dipteran species and efficient expression of the gfp marker supports the potential use of this system in other dipterans, and perhaps insects in general.

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