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题名: Selection of donor nuclei in somatic cell-mediated gene transfer using a co-transfection method
作者: Ma, Hairong; Lu, Zili; Sun, Yi; Peng, Tao; Shuai, Zhiqiang; Ma, Yufang; Zhang, Yan; Wang, Liang; Cao, Xu; Wang, Hanqing
关键词: cell cycle ; co-transfection ; mammary gland-specific promoter ; ovine adult fibroblast ; somatic cell-mediated nuclear transfer
刊名: JOURNAL OF REPRODUCTION AND DEVELOPMENT
发表日期: 2007
DOI: 10.1262/jrd.18082
卷: 53, 期:1, 页:95-104
收录类别: SCI
英文摘要: In this study, we introduced a co-transfection method for the selection of donor nuclei in somatic cell-mediated nuclear transfer. Two vectors were constructed in our experiment. One was pMSCV-GFP carrying the neomycin-resistant gene (Neo(r)) and the green fluorescent protein (GFP) reporter gene; the other was pBC1-GFP carrying the mammary gland-specific promoter and target gene GFP. Ovine adult fibroblasts were co-transfected with pMSCV-GFP and pBC1-GFP. The data from this work demonstrated that the GFP genes in both vectors could successfully co-integrate into the genomes of ovine adult fibroblasts in three of the four transgenic cell clones assayed. Furthermore, PCR analysis of transgenic embryos proved that the GFP genes in both vectors could cointegrate into the genomes of the reconstructed embryos. Subsequently, analysis of the developmental rate of the reconstructed embryos after nuclear transfer indicated that the blastocyst rate from the co-transfected donor cells was similar (approximate 8 percent) to that from individual pMSCV-GFP transfected donor cells. The influence of co-transfection resulting in modification of donor nuclei on development of reconstructed embryos was also investigated. The results of flow cytometric analysis indicated that the co-transfected ovine fibroblasts had similar quiescent characteristics in terms of cell cycle (G0+G1 percent: 73.20 +/- 4.04) to the individual pMSCV-GFP transfected fibroblasts (G0+G1 percent: 70.77 +/- 1.19) after they were treated with serum starvation for five days. Our results suggest that the co-transfection method can be used for selection of donor cell clones in somatic cell-mediated gene transfer experiments. It can be potentially extended to applications related to expression of functional protein in mammary glands and other transgenic research relevant to nuclear transfer.
WOS记录号: WOS:000244465700011
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内容类型: 期刊论文
URI标识: http://ir.xjipc.cas.cn/handle/365002/4122
Appears in Collections:资源化学研究室_期刊论文

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作者单位: Chinese Acad Sci, Xinjiang Tech Inst Phys & Chem, Urumqi 830011, Peoples R China;Chinese Acad Sci, Lanzhou Inst Chem Phys, Lanzhou 730000, Peoples R China;Xinjiang Golden Cattle Biol Incorp, Urumqi 830011, Peoples R China;Xinjiang Med Univ, Urumqi 830011, Peoples R China;Univ Alabama, Dept Pathol, Birmingham, AL 35294 USA;China Agr Univ, State Key Labs Agrobiotechnol, Coll Biol Sci, Beijing 475001, Peoples R China

Recommended Citation:
Ma, Hairong,Lu, Zili,Sun, Yi,et al. Selection of donor nuclei in somatic cell-mediated gene transfer using a co-transfection method[J]. JOURNAL OF REPRODUCTION AND DEVELOPMENT,2007,53(1):95-104.
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