The ability to chemically couple proteins to LH(N)-fragments of clostridial neurotoxins and create novel molecules with selectivity for cells other than the natural target cell of the native neurotoxin is well established. Such molecules are able to inhibit exocytosis in the target cell and have the potential to be therapeutically beneficial where secretion from a particular cell plays a causative role in a disease or medical condition. To date, these molecules have been produced by chemical coupling of the LH(N)-fragment and the targeting ligand. This is, however, not a suitable basis for producing pharmaceutical agents as the products are ill defined, difficult to control and heterogeneous. Also, the molecules described to date have targeted neuroendocrine cells that are susceptible to native neurotoxins, and therefore the benefit of creating a molecule with a novel targeting domain has been limited. In this paper, the production of a fully recombinant fusion protein from a recombinant gene encoding both the LH(N)-domain of a clostridial neurotoxin and a specific targeting domain is described, together with the ability of such recombinant fusion proteins to inhibit secretion from non-neuronal target cells. Specifically, a novel protein consisting of the LH(N)-domains of botulinum neurotoxin type C and epidermal growth factor (EGF) that is able to inhibit secretion of mucus from epithelial cells is reported. Such a molecule has the potential to prevent mucus hypersecretion in asthma and chronic obstructive pulmonary disease.

Several important groups of fungicides and insecticides are specific inhibitors of chitin synthesis in a Phycomyces enzyme system and in insect organ cultures. The recently discovered benzoylphenylurea insecticides, which prevent chitin synthesis in insect tissues, are apparently not direct-acting chitin synthetase inhibitors. These insecticides may prevent insect chitin synthesis by interfering with the proteolytic activation of the chitin synthetase zymogen.

Two cell lines were derived from primary cultures of embryonic tissue of the tobacco hornworm, Manduca sexta (L.). The cell lines were maintained on hemolymph-free synthetic insect medium. Cytogenetic and immunoligical identification of the lines were carried out. Techniques for obtaining the line and medium, subculturieng and freezing procedures for long-term storage, and the morphological and growth characteristics are described.

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