利用体细胞核移植技术生产表达红色荧光蛋白的猪转基因克隆胚胎研究

Production of Transgenic Clone Pig Embryos Expressing Red Fluorescent Protein by Somatic Nuclear Transfer Technology

[目的]为生产转基因克隆猪奠定基础。[方法]以反转录病毒转染的带有红色荧光蛋白(RFP)基因的猪胎儿成纤维细胞作为核移植的核供体,利用体细胞克隆技术研究红色荧光蛋白克隆胚胎体外发育情况。[结果]RFP转基因细胞的融合率为83.87%,与未转基因细胞(80.56%)相比无显著差异(P>0.05);RFP转基因体细胞重构胚体外囊胚率为8.67%,与未转基因细胞组(6.56%)相比无显差异著(P>0.05);RFP转基因体细胞重构胚移植于15头受体后,尚无受孕个体。[结论]利用转红色荧光蛋白基因的细胞为供体细胞,能够成功克隆出转基因胚胎,并获得转基因囊胚。

[Objective] The aim was to lay a foundation for producing transgenic clone pig.[Method] We use pig fetus fibroblasts with red fluorescent protein（RFP） that transfect with retrovirus for donor of nuclear transfer,utilize somatic cell cloning technology to research the development in vitro of cloning embry expressing red fluorescent protein.[Result]The results indicated that fusion rate of RFP transgene cell and the control are 83.87% and 80.56% respectively,without significant difference（P〉0.05）;blastula rate in vitro of RFP transgene cell reconstructed embryo is 8.67% compare to the control（6.56%） without significant difference（P〉0.05）;RFP transgene cell reconstructed embryo transfer to fifteen surrogacy and none conception.[Conclusion]Use of red fluorescent protein transgene cell for donor cells can successfully clone transgenic embryo,obtain transgenic blastula.