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通过体细胞核移植技术高效生产 ω-3 脂肪酸去饱和酶(sFat-1)转基因猪。

Efficient production of omega-3 fatty acid desaturase (sFat-1)-transgenic pigs by somatic cell nuclear transfer.

机构信息

Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, China.

出版信息

Sci China Life Sci. 2010 Apr;53(4):517-23. doi: 10.1007/s11427-010-0080-x. Epub 2010 May 7.

Abstract

Omega-3(omega-3) fatty acid desaturase transgenic pigs may improve carcass fatty acid composition. The use of transgenic pigs is also an excellent large animal model for studying the role of omega-3 fatty acids in the prevention and treatment of coronary heart disease and cancer. Transgenic pigs carrying synthesized fatty acid desaturase-1 gene (sFat-1) from Caenorhabditis briggsae by somatic cell nuclear transfer (SCNT) were produced for the first time in China. Porcine fetal fibroblast cells were transfected with a sFat-1 expression cassette by the liposome-mediated method. Transgenic embryos were reconstructed by nuclear transfer of positive cells into enucleated in vitro matured oocytes. A total of 1889 reconstructed embryos were transferred into 10 naturally cycling gilts. Nine early pregnancies were established, 7 of which went to term. Twenty-one piglets were born. The cloning efficiency was 1.1% (born piglets/transferred embryos). The integration of the sFat-1 gene was confirmed in 15 live cloned piglets by PCR and Southern blot except for 2 piglets. Expression of the sFat-1 gene in 12 of 13 piglets was detected with RT-PCR. The data demonstrates that an efficient system for sFat-1 transgenic cloned pigs was developed, which led to the successful production of piglets expressing the sFat-1 gene.

摘要

ω-3 脂肪酸去饱和酶转基因猪可能改善胴体脂肪酸组成。转基因猪也是研究 ω-3 脂肪酸在预防和治疗冠心病和癌症中的作用的极好的大型动物模型。通过体细胞核移植 (SCNT),首次在中国生产出携带合成脂肪酸去饱和酶-1 基因 (sFat-1) 的转基因猪。通过脂质体介导的方法将 sFat-1 表达盒转染到猪胎儿成纤维细胞中。将阳性细胞的核转移到体外成熟去核卵母细胞中,构建转基因胚胎。总共将 1889 个重构胚胎移植到 10 头自然循环的小母猪中。建立了 9 例早期妊娠,其中 7 例足月。出生了 21 头小猪。克隆效率为 1.1%(出生仔猪/移植胚胎)。除了 2 头小猪外,通过 PCR 和 Southern blot 证实了 15 头活体克隆仔猪中 sFat-1 基因的整合。在 13 头仔猪中的 12 头中通过 RT-PCR 检测到 sFat-1 基因的表达。数据表明,开发了一种有效的 sFat-1 转基因克隆猪系统,成功生产了表达 sFat-1 基因的仔猪。

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