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Transgene expression of enhanced green fluorescent protein in cloned rabbits generated from in vitro-transfected adult fibroblasts

Transgene expression of enhanced green fluorescent protein i...

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第九届中国实验动物科学年会（2010新疆）

作者： shangang li yi guo jianjun shi chunguang yin fengying xing lingyang xu chuanshan zhang tao liu yao li hongbin li lixin du xuejin chen

Objective live rabbits have previously been generatedthrough nuclear transfer using adult cells as nuclear *** demonstrated in this study that transfected adult rabbit fibroblasts are also capableof supporting full-term *** fibroblasts were transfected with a pEGFP-C1 plasmid using lipofectamine TM 2000,and the transgenic cells were derived from conditioned *** transgenic fibroblasts were cultured until confluent and then serum-starved prior to be used as nuclear *** nuclear transfer and activation,22%(12/55) of the transgenic cloned embryos developed to the blastocyst stage.A total of 114 embryos at the 4- to 8-cell stage were transferred to the oviducts of 8 pseudo-pregnant mothers;5 of these animals became pregnant,and 3 of the 5 mother rabbits carried the pregnancy to term. Caesarean section was performed on the 3 pregnant mothers,yielding 4 kits,one of which has survived for more than 9 *** fluorescence could be detected in the toenails of the living cloned rabbit and the offspring from the living cloned rabbit under ultraviolet *** analyses confirmed that all 4 cloned rabbits were genetically identical to the transgenic donor cells,and that they all carried the EGFP *** present study demonstrated that transgenic rabbits can be generated through nuclear *** results may facilitate future developments in the genetic engineering of rabbits.

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