Germline transformation of Drosophila melanogaster with the piggyBac transposon vector

作者：Alfred M. Handler, Robert A. Harrell · 发表于：Insect Molecular Biology · 年份：1999 · DOI：10.1046/j.1365-2583.1999.00139.x · 被引用次数：280 · 研究领域：CRISPR and Genetic Engineering、Insect Resistance and Genetics、Animal Genetics and Reproduction

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.