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作 者:杭苏琴[1] 刘为敏[1] 牛树理[1] 冯紫云[1]
机构地区:[1]南京农业大学动物科技学院,江苏南京210095
出 处:《畜牧与兽医》2003年第12期3-4,共2页Animal Husbandry & Veterinary Medicine
摘 要:BALB/C雄性小鼠脾细胞悬液腹腔注射免疫同系母鼠 ,精子细胞毒性试验筛选抗血清 ,效价高的抗血清分别用于胚胎培养的毒性试验和制备单克隆抗体再辅以间接免疫荧光和PCR对胚胎进行性别鉴别。结果表明 ,直接用H Y抗血清培养的胚胎 ,胚胎退化率达 43 3 % ,与自然性比差异不显著 ;PCR验证间接免疫荧光法鉴定的胚胎性别 ,雄性胚胎准确率为 83 % ,雌性胚胎准确率为 94%。BALB/C female mice were immunized by intraperitoneal injections of syngeneic male spleen cells and antiserum was detected by sperm cytotoxicity test.Then the chosen antisera were used in mouse embryos culture in vitro and to prepare H-Y monoclonal antibody for later sex detection by indirect immuno-fluorescence assay and polymerase chain reaction(PCR)with amplification of Y chromosome specific sequence.Cultured embryos were classified either affected or unaffected,affected embryos was 43.3% without significant difference from the normal sex ratio.The results from PCR and indirect immuno-fluorescence assay showed that male embryos of H-Y positive was 83%(15/18),female embryos of H-Y negative was 94%(15/16).
关 键 词:雄性特异性抗原 H-Y抗体 小鼠胚胎 性别鉴定 单克隆抗体 脾细胞注射 精子细胞毒性试验
分 类 号:S852.4[农业科学—基础兽医学] S814.7[农业科学—兽医学]
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