MOLECULAR BIOLOGY Pink Bollworm Larvae Infection with a Double Subgenomic Sindbis (dsSIN) Virus to Express Genes of Interest

Pest control strategies that rely on genetic manipulation of pest insects depend on proper expression of foreign genes in the pest, as well as on experimental evaluation of genes introduced by transformation with various chimeric fusion constructs, following the work of Thomas et al. (2000). The evaluation of such constructs relies heavily on detection of gene products and demonstration of their effects in vivo. A dependable genetic expression system that reliably can direct production of heterologous or engineered gene products at high levels would be of significant value to establish the presence and activity of the foreign proteins that are to be encoded in these constructs. It is also helpful to elucidate the biological effects of any engineered proteins in intact organisms. To this end, we report the expression of the heterologous gene, green fluorescent protein, from a viral vector in a major lepidopteran pest of cotton. The double subgenomic expression vector in this study was based upon the Sindbis (SIN) alphavirus. These dsSIN viruses have been used to express a spectrum of genes: chloramphenicol acetyltransferase (Olson et al., 1994; Kamrud et al., 1995; Kamrud et al., 1997); luciferase (Johnson et al., 1999), a toxin gene from Androctonus australis Hector (Higgs et al., 1995a); green fluorescent protein from Aequore