Gao Shaorong
Fels Institute for Cancer Research and Molecular Biology, Temple University School of Medicine, Philadelphia, PA, USA.
Methods Mol Biol. 2006;325:25-33. doi: 10.1385/1-59745-005-7:25.
Cloning by nuclear transfer in mammals has revealed the remarkable ability of an oocyte to reprogram transferred cell nuclei and induce them to recapitulate the developmental program. This chapter summarizes the method used since 1998 for mouse cloning, which differs from that for large animal cloning. A Piezo-drill micromanipulator allows direct injection of nuclei into enucleated mice oocytes instead of the electrofusion method used for introducing a nucleus into an enucleated oocyte of a large animal. After activation, reconstructed embryos are allowed to develop to the morulae or blastocyst stage before transfer into surrogate mothers.
哺乳动物的核移植克隆技术揭示了卵母细胞对移植细胞核进行重编程并诱导其重现发育程序的非凡能力。本章总结了自1998年以来用于小鼠克隆的方法,该方法与大型动物克隆方法不同。压电钻显微操作器可将细胞核直接注入去核的小鼠卵母细胞,而不是用于将细胞核导入大型动物去核卵母细胞的电融合方法。激活后,将重构胚胎培养至桑葚胚或囊胚阶段,再移植到代孕母体中。