The Immunogenicity And Fertilization Of The Recombinant Proteins TMDC-Ⅲ,SP-10,AEG

Fertilization is a highly programmed process by which two radically different cells, sperm and egg, unite to form a zygoL It is mediated by complementary molecules present on the surface of the respective gametes. Especially the proteins in the sperm acrom play an important role, but only a little can be extracted from sperm. Now large amount of proteins can be obtained easily using biological technique. In this experiment three sperm proeins, SP-10.AEG.tMDC-III.that were cloned and expressed by Brist U.K., immunized on rats for analysing their immunogenicity and the effects on the fertility. Then using indirect immuno-fluorescein staining method, the localization of SP-10, tMDC-III and AEG on human, mouse, rabbit, pig, bull sperm was observed under confocomicroscopy.The results as follows:1 .The rats successfully produced antibody after immunization with antigens SP-10, AEG, tMDC-III respective. Indirect immunofluorescence and ELISA studies proved the three proteins maintained their native nature of immunogenicity. They can be used in further studies.2. After immunization mating experiment was conducted,the results shown fertility rate are 91.6%,83.3%,75.0% and 58.3% in control, tMD III, Sp-10 and AGE group respectively. The above results shown immunized AGE reduced the fertility rate.3. Testis, uterus, ovary, epididymis and fallopian tube were fixed and cutted for pathology examination, we have not found significantly change on testis, uterus, ovary, epididymis and fallopian tube compared with control. It suggested that the antibody did not inhibit sperm development and damage the reproductive tract to decrease the fertility rate. It also indicated the antigens used in this experiment are not lexicological for the rats, supporting the use of them for contraceptive vaccine.4. Using indirect immuno-fluorescein staining method, the localization of Sp-10, tMDC-III and AEG on human, mouse, rabbit, pig, bull sperm was observed under confocomicroscopy. It was found that antigens Sp-10 and AEG express on acrosome of all kinds sperms, supporting their roles in sperm-egg fusion. It was firstly described that tMDC-III express mostly on the equator, neck and some part of tail of human sperm, on the acrosome of mouse and bull sperms, on the acrosome and neck of pig sperm, on the head, neck, and middle piece, principal piece of tail of rabbit sperm. It implied tMDC-III may associate with sperm-egg fusion.Master: Wang Feng-chao Major: Basic Veterinary Supervisor: Hao \kn-hong Li Qing-zhang...