SYNTHETIC PHEROMONE BIOSYNTHESIS ACTIVATING NEUROPEPTIDE GENE EXPRESSED IN A BACULOVIRUS EXPRESSION SYSTEM

A synthetic gene of the pheromone biosynthesis activating neuropeptide (PBAN) of corn earworm Helicoverpa tea, with and without a signal seq uence of the cuticle protein of Drosophila melanogaster, was cloned be hind the polyhedrin promoter of AcMNPV. Two recombinant baculoviruses were constructed and used to infect a number of insect cell lines incl uding Sf9 and 5B1-4. High pheromonotropic activity was consistently ob tained from 5B1-4 cell culture that was infected with the recombinant baculovirus vLNV-4 containing the signal sequence. The PBAN gene-produ ct was isolated by HPLC and analyzed by electrospray ionization mass s pectrometry. Low levels of biological activity obtained from Sf9 cells infected with the recombinant virus vPBAN may be due to lack of prope r amidation at the C-terminus of the expressed peptide or rapid proteo lytic degradation of the product.