pcxn2mizumo对小鼠c57bl6生育能力的影响-effects of pcx n2 m izumo on the fertility of c57bl6 mice.docxVIP

说明对照质粒构建成功，重组质粒正确无误；②RT-PCR 用 1％的琼脂糖凝胶电泳得到RT-PCR 产物为 468 bp，产物碱基测序结果经 BLAST 比对分析后证实为 C57BL/6 小鼠 Izumo 的特异性 mRNA 片断，说明质粒 pCXN2-mIZUMO 在小鼠的体内可以表达；③ELISA 检测 重组质粒 pCXN2-mIZUMO 第一次免疫小鼠后 2 周即可检测到抗 Izumo 特异性抗 体，经过二次加强免疫后抗体水平从 0~6 周持续升高并维持在较高的水平；④体外受精试 验 实验组受精率为 11.6%，对照组受精率为 25.2％，统计分析表明差异有统计学意义， P0.05；⑤实验动物合笼 免疫雌鼠与正常雄鼠交配，实验组与对照组间产仔数的差异具 有统计学意义，P0.05；而免疫雄鼠与正常雌鼠交配，实验组与对照组的产仔数无统计学 差异。【结论】①对照质粒 pCXN2 构建成功；②重组质粒 pCXN2-mIZUMO 可以在小鼠骨骼 肌细胞中表达；③经重组质粒 pCXN2-mIZUMO 免疫的小鼠体内诱发了抗 Izumo 特异性免 疫应答。；④经重组质粒 pCXN2-mIZUMO 免疫后的小鼠抗血清在体外可以降低其受精率；⑤经重组质粒 pCXN2-mIZUMO 免疫的雌性小鼠的生育能力下降；⑥为 Izumo 作为基因免 疫避孕的靶抗原研究提供了新的实验证据。【关键词】Izumo；免疫避孕； 基因免疫； C57BL/6 小鼠Effect of recombinant plasmid pCXN2-mIZUMO on the Fertility of C57BL/6 MicePostgraduate: GangAN Supervisor: Prof. Tian-huaHUANGABSTRACT[BACKGROUND AIM] It is a very interesting project for scientists to explore a method of convenience and availability for human family planning and pest animal birth control. Their work focuses on application of immunocontraception vaccine in many countries, especially developing ones. In the developed world, steroid contraceptives for humans are both widely used and efficacious. Elsewhere they are too costly. The development of less expensive methods is considered necessary. One such method is immunocontraception, i.e. the vaccination against sperm, egg, or reproductive hormones to prevent either fertilization or the production of gametes. With the development of immunology and gene engineering, gene immunization, also known as DNA or polynucleotide immunization, is becoming a novel strategy for vaccine development in which plasmid DNA encoding either individual or a collection of antigens is directly administered to a host. Such immunization leads to host expression of the delivered foreign gene, resulting in the induction of a specific immune response against the in vivo produced antigen. In March 2005 Naokazu Inoue et al firstly reported a novel protein named Izumo which belongs to immunoglobulin superfamily. Izumo