The lepidopteran transposon vector, piggyBac, mediates germ-line transformation in the Mediterranean fruit fly.
The piggyBac (IFP2) short inverted terminal repeat transposable element from the cabbage looper Trichoplusia ni was tested for gene transfer vector function as part of a bipartite vector-helper system in the Mediterranean fruit fly Ceratitis capitata. A piggyBac vector marked with the medfly white gene was tested with a normally regulated piggyBac transposase helper at two different concentrations in a white eye host strain. Both experiments yielded transformants at an approximate frequency of 3-5%, with a total of six lines isolated having pigmented eyes with various levels of coloration. G1 transformant siblings from each line shared at least one common integration, with several sublines having an additional second integration. For the first transformant line isolated, two integrations were determined to be stable for 15 generations. For five of the lines, a piggyBac-mediated transposition was verified by sequencing the insertion site junctions isolated by inverse PCR that identified a characteristic piggyBac TTAA target site duplication. The efficient and stable transformation of the medfly with a lepidopteran vector represents transposon function over a relatively large evolutionary distance and suggests that the piggyBac system will be functional in a broad range of insects.
Handler
A M
AM
Center for Medical, Agricultural, and Veterinary Entomology, Agricultural Research Service, U.S. Department of Agriculture, Gainesville, FL, 32608, USA. handler@nervm.nerdc.ufl.edu
McCombs
S D
SD
Fraser
M J
MJ
Saul
S H
SH
D000818:Animals; D001483:Base Sequence; D004251:DNA Transposable Elements; D004175:Diptera / Q000235:genetics*; D005260:Female; D005822:Genetic Vectors; D018095:Germ-Line Mutation; D007915:Lepidoptera / Q000235:genetics*; D008297:Male; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D009693:Nucleic Acid Hybridization; D017422:Sequence Analysis, DNA
D004251:DNA Transposable Elements