The Tissue Distribution Of Serine Proteinase Inhibitor 1 And Prokaryotic Expression Of Storage Protein From Ostrinia Furnacalis Guenee Larvae

Serine protease inhibitors (Serpins) are a super family of proteins, most of which control protease-mediated processes and are evolved to the regulation of serine protease (SPs) to maintain homeostasis. So far, over 800 Serpins have been identified in animals, plants, bacteria, and viruses. In order to explore Serpinl in insect immune function, we have obtained Serpinl antibody, using indirect immunofluorescence labeling and Western blot to detect Serpinl in Ostrinia furnacalis larvae, to analyze of immune regulation at the protein level, and to study the mechanismof serpinl on the control of PPO activating system, Which is helpful to find the protein enzyme activation pathway in insect development and immune response. The results showed that can find that Serpinl in various kinds of hemocytes, such as, granular hemocytes, plasmatocytes and oenocytoids, presenting green fluorescent under the fluorescence microscopy with indirect immunofluorescence labeling. According to the density of protein strips, Western-blot analysis was used to identify Serpinl expression the amount. The result revealed that the highest level was in the integument, and then the hemolymph and midgut, and the lowest level was found in fat body. According to the gold particles numbers in different ten microscopic fields, colloidal gold marked immune electron microscopy experiments was used to identify Serpinl expression the amount. The result showed that the highest level was in the integument, and then the hemolymph and midgut, and the lowest level was found in fat body.Insect storage protein (SP) is composed of six subunit protein aggregation of spherical six polymers. It has great homology with hemocyanin. SP is a protein that insects stored as nutrients for later use, and it in mainly fluid protein is a complete metamorphosis insect larvae. It is important for adult normal development and the female egg as the amino acids repository. Meanwhile, SP may be involved in the insect immunity. In order to further study on storage protein, SP gene was cloned from 5th instars Ostrinia furnacalis larvae and was used to construct a pET-30a-SP prokaryotic expression vector, and expression condition for the recombinant O. furnacalis SP was optimized in the in Escherichia coli.