不同类型的转基因细胞核供体对生产小鼠转基因克隆胚胎的影响

闫晓飞;徐燕宁;李洪武;关娜;单智焱;钟淑琦;金连弘;雷蕾 

解剖学报 ›› 2009, Vol. 40 ›› Issue (3) : 409-413.

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解剖学报 ›› 2009, Vol. 40 ›› Issue (3) : 409-413. DOI: 10.3969/j.issn.0529-1356.2009.03.013
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不同类型的转基因细胞核供体对生产小鼠转基因克隆胚胎的影响

  • 闫晓飞1 ;徐燕宁1; 李洪武2 ;关娜1; 单智焱1 ;钟淑琦1; 金连弘1 ;雷蕾1* 
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Production of transgenic cloned mouse embryos by somatic cell nuclear transfer with different donor cells

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Abstract

Objective To generate transgenic mouse embryos by somatic cell nuclear transfer (SCNT), and compare effects of different donor cells on developmental potential of cloned embryos. Methods Mouse embryo fibroblasts and mouse embryonic stem (ES) cells were transfected with a plasmid (pCE-EGFP-IRES-NEO) containing the enhanced green fluorescent protein (GFP) and neomycinresistant (Neor) genes by electroporation. Transgenic cell lines were obtained for SCNT after 14 days selection with G418. At the same time, SCNT was performed using cumulus cells and normal fibroblasts as control. Results There was no significant difference in blastocyst rates between reconstructed embryos derived from transgenic fibroblasts and nontransgenic ones (154/160,19.23% vs 22.91%, EM>P/EM>>0.05). Reconstructed embryos from transgenic ES cells showed higher blastocyst developmental rates than that from transgenic fibroblasts donor (152/154,41.54% vs 19.23%, P<0.05). Cumulus cloned embryos had better developmental potential than that of fibroblasts cloned ones (171/160, 41.17% vs 22.91%, P<0.05). Conclusion Our results showed that transgenic donor cells did not affect EM>in vitro/EM> developmental potential of mouse SCNT embryos. Using EGFP maker, mouse transgenic blastocysts could be produced effectively

关键词

核移植 / 转基因体细胞 / 电穿孔 / 绿色荧光蛋白 / 小鼠

Key words

Nuclear transfer / Transgenic somatic cell / Electroporation / green flourescent protein / Mouse

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闫晓飞;徐燕宁;李洪武;关娜;单智焱;钟淑琦;金连弘;雷蕾 . 不同类型的转基因细胞核供体对生产小鼠转基因克隆胚胎的影响[J]. 解剖学报. 2009, 40(3): 409-413 https://doi.org/10.3969/j.issn.0529-1356.2009.03.013
Production of transgenic cloned mouse embryos by somatic cell nuclear transfer with different donor cells[J]. Acta Anatomica Sinica. 2009, 40(3): 409-413 https://doi.org/10.3969/j.issn.0529-1356.2009.03.013
中图分类号: R329.1   

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