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猪体细胞核移植及几个相关问题的研究论文.pdf62页
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东北农业大学 博士学位论文
猪体细胞核移植及几个相关问题的研究 姓名:孙兴参 申请学位级别:博士 专业:动物组织胚胎学 指导教师:谭景和
..塾!丝 塑
里 .!塑!:!一 摘 要 细胞核移植技术不仅是研究细胞核与细胞质相互作用的重要手段,而且是在医学和
农业等领域都将有着广泛的应用前景的一项重要胚胎工程技术。尽管近来哺乳动物体细
胞核移植克隆已经取得了很大进展,但实践证明猪的克隆难度最大,有许多问题亟待解
决。本实验对猪体细胞核移植及其相关问题如细胞融合、卵母细胞激活和卵母细胞体外
成熟进行了研究。结果如下: 1.将卵丘细胞直接注入卵母细胞或将培养的颗粒细胞与卵母细胞融合,供核细胞都能
发生染色质提前凝集 Pcc :将培养的颗粒细胞与去核的卵母细胞融合后,核重组变化
方式与直接注射法的结果相似,但是融合培养的颗粒细胞要比直接注射卵丘细胞的发育
速度快。 2.将卵丘细胞直接注入卵母细胞构建核移植胚胎,在中间受体内发育到囊胚。 3.猪.猪同种核移植胚胎和猪.小鼠异种移核胚胎激活后,异种重组胚多形成二原核 2PN ,而同种重组胚则主要形成一原核 IPN 。 %和88.9%:电刺激屉DMAP与化学刺激,6.DMAP激活方法在激活率上虽然没有明显差
异,但电激活的卵裂率明显高于化学激活。 5.采用相对较低场强 0.8v/cm左右 和长脉冲 60杠s 组合的重组胚胎的融合率
高于高场强 2.1kv,cm左右 和短脉冲 20us 的组合。 6.采用活体荧光染料Hoc∞t33342对猪的生发泡进行了观察,发现猪Gv染色质构型
有五种类型:腔前卵泡卵母细胞都为Gv.o染色质构型;随着卵泡的发育,Gv一1卵母细
胞比例增高;体外成熟过程中,随
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参考资料

 

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