Production of transgenic calves by somatic cell nuclear transfer
Production of transgenic calves by somatic cell nuclear transfer作者机构：StateKeyLaboratoryforAgrobiotechnologyChinaAgriculturalUniversityBeijing100094China InstituteofAnimalScienceChineseAcademyofAgriculturalScienceBeijing100094China GentitanBiotechnologyLtd.Beijing100084China
出 版 物：《Chinese Science Bulletin》 (中国科学通报)
年 卷 期：2004年第49卷第2期
页 面：161-166页
核心收录：
学科分类：0710[理学-生物学] 07[理学] 08[工学] 09[农学] 071007[理学-遗传学] 0901[农学-作物学] 0836[工学-生物工程] 090102[农学-作物遗传育种]
基 金：State “863” High-Tech Research and Development Project, (2001AA213091, 2002AA206111) Natural Science Foundation of Beijing Municipality, (5030001) Natural Science Foundation of Beijing Municipality
主 题：核移植 体细胞 增强绿荧光蛋白质 EGFP 转基因技术
摘 要：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.
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