GREEN FLUORESCENT PROTEIN EXPRESSED IN LIVING MOSQUITOS - WITHOUT THEREQUIREMENT OF TRANSFORMATION

Mosquitoes transmit viruses, protozoa and nematodes that are major cau ses of morbidity and mortality in humans. Details of arthropod anatomy and development, and the replication and development of pathogens in the arthropod vector have relied upon examination of dissected or hist ologically processed material. We constructed a double-subgenomic Sind bis (dsSIN) virus expressing green fluorescent protein to demonstrate the potential of this protein for studying pathogen development in liv ing arthropods. We were able to observe dissemination of virus, and fu rthermore, it was possible to observe components of the nervous system of mosquito larvae in extraordinary detail and record this on video t ape. Although green fluorescent protein has been used as a reporter ge ne in a number of organisms, expression has relied upon transformation of cells or embryos. Transformation technology has limited applicabil ity, thus we have described an alternative system that, due to the bro ad host range and viral tropisms of dsSIN viruses, may be useful to sc ientists in a range of disciplines. Green fluorescent protein may also provide a nonlethal selection method for use in transgenic arthropod research.