Efficient production of omega-3 fatty acid desaturase (sFat-1)-transgenic pigs by somatic cell nuclear transfer
Efficient production of omega-3 fatty acid desaturase (sFat-1)-transgenic pigs by somatic cell nuclear transfer
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DOI：
10.1007/s11427-010-0080-x
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发表时间：
2010-04-01
影响因子：
9.1
通讯作者：
Chen HongXing
中科院分区：
文献类型：
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作者：
Pan DengKe;Zhang Li;Chen HongXing
转omega-3(omega-3)脂肪酸去饱和酶基因猪可改善屠体脂肪酸组成。转基因猪的使用也是研究omega-3脂肪酸在预防和治疗冠心病和癌症中的作用的优秀大型动物模型。利用体细胞核移植技术，在中国首次获得了携带桥小杆线虫合成脂肪酸去饱和酶-1基因的转基因猪。用脂质体介导法将SFAT-1表达盒导入猪胎儿成纤维细胞。通过将阳性细胞核移植到体外去核的成熟卵母细胞中重建转基因胚胎。将1889枚重组胚胎移植到10头自然循环母猪体内。已确定9例早孕，其中7例足月妊娠。21头小猪出生了。克隆效率为1.1%(初生仔猪/移植胚胎)。经PCR和Southern印迹分析，除2头外，其余15头活体克隆仔猪均整合了SFAT-1基因。用RT-PCR方法检测了13头仔猪中12头的SFAT-1基因的表达。结果表明，建立了一套高效的SFAT-1转基因克隆猪系统，成功地培育出了表达SFAT-1基因的仔猪。
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.