Insect Molecular Biology (2002) 11(4), 291–297 © 2002 The Royal Entomological Society 291 Blackwell Science, LtdGerm-line transformation of the South American malaria

Stable and efficient germ-line transformation was achieved in the South American malaria vector, Anoph-eles albimanus, using a piggyBac vector marked with an enhanced green fluorescent protein gene regulated by the Drosophila melanogaster polyubiquitin promoter. Transgenic mosquitoes were identified from four independent experiments at frequencies ranging from 20 to 43 % per fertile G0. Fluorescence was observable throughout the body of larvae and pupae, and abdom-inal segments of adults. Transgenic lines analysed by Southern hybridization had one to six germ-line inte-grations, with most lines having three or more integra-tions. Hybridized transposon vector fragments and insertion site sequences were consistent with precise piggyBac-mediated integrations, although this was not verified for all lines. The piggyBac/PUbnlsEGFP vector appears to be a robust transformation system for this anopheline species, in contrast to the use of a piggy-Bac vector in An. gambiae. Further tests are needed to determine if differences in anopheline transformation efficiency are due to the marker systems or to organis-mal or cellular factors specific to the species