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Production of transgenic calves by somatic cell nuclear transfer

Chinese Science Bulletin 2004年第2期49卷 161-166页

作者： gongguochun DAIYunping FANBaoliang ZHUHuabing WANGLili WANGHaiping TANGBo LIUYing LIRong WANGRong HUANGYinghua LINing StateKeyLaboratoryforAgrobiotechnology ChinaAgriculturalUniversityBeijing100094China InstituteofAnimalScience ChineseAcademyofAgriculturalScienceBeijing100094China GentitanBiotechnologyLtd. Beijing100084China

Bovine fetal oviduct epithelial cells were transfected with constructed 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 transgenic cell line was obtained. Somatic cell nuclear transfer (SCNT) was cartied 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 *** and Southern blot analysis confirmed the integration of transgene in all 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 be ensured to be transgenic by efficiently pre-screening transgenic cells and SCNT embryos using the constructed double marker selective vector.

关键词：核移植体细胞增强绿荧光蛋白质 EGFP 转基因技术

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