黄牛和水牛种间嵌合的初步研究-动物遗传育种与繁殖专业毕业论文.docxVIP

硕士论文：黄牛和水牛静闻嵌合的翅步研究黄牛和水牛种闻嵌合的初步研究 硕士论文：黄牛和水牛静闻嵌合的翅步研究 黄牛和水牛种闻嵌合的初步研究 摘要 本研究系统探讨了影响黄牛和水牛嵌合体制作的主要因素，初步建立了一套黄 牛和承牟稀简散合的獠净与方法。 注入嵌合法采用机械剥离法或免痰外科法分离胚胎内细胞团(IcM)，然后注 入骜邑去除la蛭豹受俸囊籁巾构建承牟和黄牛静嵌会珏：聚合袋舍法采矮链酶蛋 白酶消化透明带或用机械剥离法去除潦明带，然后在禽有100“g／ml PHA的培养液 孛聚合影或嵌会莲。＼缭寨发瑗，在爱免疫岁}萃摹法分离lC醚辩，抗蠢漓懿灭活温度 从57℃升至63．5℃，lCM的获得数显藩升高(0％vs 100％，P0．01)，如蓿在分 褰培葵渡中添麓6％夔熬犊盘漆(KS)，ICM静获褥数大大黪低(97。6％VS穆％， P0．01)。采用免疫外科法分离得到的黄牛lCM进行水牛囊胚的IcM置换熏组， 其重缀疆豹存滔率与搬壤裂蹇法褥裂瓣ICM无显藩差异(91。4％vs 87+S％， P0．05)；但囊胚孵化率则显著提高(80％vs 43．8％；P0．05)。8-细胞黄牛胚胎聚 会水牛爨堪驱岛黄牛囊憝聚合水牛桑攥坯甥比，聚合疑存话零(ss。7％vs 77．8％) 和囊瓤发育率(72．2％vs 42．9％)均无湿著差弊(P0．05)。采用显微手术法分离黄 牛和水牛8-细胞胚胎卵裂球进行聚会，聚合率为92．3％，囊胚发育率为58．3％，与 用0．25％链酶凝自酶分离胚胎卵袭球进行胚胎聚合的聚合率(86．7％)和囊胍发育 率(46．2％)均无显著差异(po．05)。 黻～滞采表明：(1)承牛帮黄牛胚狳通过郛袈球聚含或ICM置换获得的耱闻嵌 合胚胎惩继续发育；(2)用于黄牛ICM分离的兔抗牛抗衄清需在63．5。C灭活30min， 置分离嚣在无纛涛懿壤养滚串进行；《匐透过分离ICM譬换遴行惩麓嵌合爵，免疫 外科法优于显微手术法；(4)胚胎聚合前的发育阶段对其聚合成功率和随后的胚胎 发弯秃缓嚣影穗。(静嚣毅雾裂稼酶分离方法(显徽手零法秘辩漪纯法)对荚聚合 率和囊胚发育翠无明显影响。 关键词：黄牛水牛内细胞团艘换，胚胎聚合、嵌合体 。 { ’ 堕主笙苎!堕生塑查生登塑堂全竺塑生里塞 堕主笙苎!堕生塑查生登塑堂全竺塑生里塞 一 A PRELIMINARY STUDY ON EMBRYONIC CHIMERAS BETWEEN BUFFALo AND CATTLE ABSTRACT This study was undertaken to systematically examine factors affecting the chimeric efficiency of buffalo and bovine embryos，so as to establish a preliminary procedure for making interspecific chimeras between cattle and bu助lo． Bovineinnercellmass(ICM)wasisolatedfromblastocystsby eithermechanicalmethod or immunosurgery,and then injected into buffalo blastocysts that ICM was removed to reconstruct a chimeric embryo．In addition，chimeric embryos were also reconstructed by aggregating blastomeres from early bovine and buffalo embryos isolated by either enzymatic or mechanical method in a droplet of culture medium containing 1001xg／ml phytohemagglutinin(PHA)． Significantly more ICM were isolated from bovine blastocysts by immunosurgery when the anti．cattle rabbit serum was inactivated at 63．5。C in comparison with him inactivated at 56。C (100％vs．O％，PO．01)．Supplementing isolating medium with 6％FCS resulted in a significant decrease in