Effects Of Double Inhibin Gene Immunization On Mouse Reproductive Hormone

There was high homogeneity of physical and chemical swine ovarian inhibin with human, bovine, ovine inhibin and other protein in the body. The fused protein has high homology with other proteins in the NR protein bank, so it is not suitable for inhibin antigen. The results indicated that α (1³2) and α (1³2) regions were two antigenic determinants of inhibin. So gene encoding inhibin α (1-32) is an candidate one for construct inhibin gene vaccine. Doubled gene encoding porcine inhibin α (1 — 32) was cloned and recombined with pcDNA3.1 expression vector to construct inhibin gene vaccine pcINH. The recombinant plasmid pcINH was expressed in E. coli. DH5 α .The expression protein had inhibin immunization activity and has no homology with other protein in the body.Plasmid pcINH were injected to mouse skeletal muscle at 0μ.g, 0.5μg, 1.0μg, 1.5μg and 2.0μg in each group.The first boost was conducted after 15d of the primary boost. The plasma were prepared before the primary boost, first boost and 15d after the first boost and stored at -20°C. Inhibin antibody of the plasma sample was assayed with ELISA. There is a remarkable difference in plasma FSH between positive and control mouses on day 15 after primary immunization of pcINH (p<0.05). After immunization of pcINH, the level of FSH in positive mouses was higher than that in negative mousse (p>0.05). The level of plasma LH went up after twice and three times immunization of pcINH, but there was no difference between positive and control mousse (p>0.05). The plasma E₂ went up after primary immunizations of pcINH (p>0.05). It went up remarkably on day 15d after booster injection of pcINH (p<0.05) and rose fatherly after three immunizations of pcINH (p<0.01). The plasma LH went up after twice and three times immunizations of pcINH. After three times immunizations, 1.0 μg pcINH promoted significantly high level of LH, but there was no difference between different dosage groups (p>0.05). The level of E₂ went up after twice and three times immunization of pcINH, but there was no difference between positive and negative mousse (p>0.05). In conclusion, inhibin gene immunization can stimulate follicular development of mouses. The increase of booster times can promote the maturation of follicular. Inhibin gene immunization can raise the level of plasma FSH, E₂ and LH, which set up the basis of inhibin gene immunizations on large animals. In order to improve animal reproductive traits using inhibin gene immunization, these suggestions should be considered for the ne,xt experiment, optimizing immunization methods, such as immunization time, dose, injection method and boost.