Production of transgenic calves by somatic cell nuclear transfer

文献类型： 外文期刊

第一作者： Gong, GC

作者： Gong, GC;Dai, YP;Fan, BL;Zhu, HB;Wang, LL;Wang, HP;Tang, B;Liu, Y;Li, R;Wan, R;Huang, YH;Li, N

作者机构：

关键词： transgenic;somatic cell;nuclear transfer;EGFP;bovine

期刊名称：CHINESE SCIENCE BULLETIN （ 影响因子：1.649； 五年影响因子：1.738 ）

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收录情况： SCI

摘要： Bovine fetal oviduct epithelial cells were transfected with construeted double marker selective vector (pCE-EGFP-IRES-Neo-dNdB) containing the enhanced green fluorescent protein (EGFP) and neomycin-resistant (Neo~r) genes by electroporation, and a transgenie cell line was obtained. Somatic cell nuclear transfer (SCNT) was carried out using the transgenic cells as nuclei donor. A total of 424 SCNT embryos were reconstructed and 208 (49.1%) of them developed to blastocyst stage. 17 blastocysts on D 7 after reconstruction were transferred to 17 surrogate calves, and 5 (29.4%) recipients were found to be pregnant. Three of them maintained to term and delivered three cloned calves. PCR and Southern blot analysis eonfirmed the integration of transgene inall of the three cloned calves. In addition, expression of EGFP was detected in biopsy isolated from the transgenic cloned calves and fibroblasts derived from the biopsy. Our results suggest that transgenic calves could be efficiently produced by SCNT using transgenic cells as nuclei donor. Furthermore, all cloned animals could he ensured to be transgenic by efficiently prc-screening transgenic cells and SCNT embryos using the constructed double marker selective vector.

分类号： N1

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