The production of transgenic pigs for xenotransplantation

1)转基因猪生产的基本技术，如卵母细胞的体外成熟、受精和培养、核移植、胚胎冷冻、胚胎移植、2）丝裂素活化蛋白激酶（mitosen-activated protein kinase，MAPK）介导成熟诱导信号从细胞质进入细胞核，诱导猪卵母细胞减数分裂重新启动; 3）透明质酸对猪卵母细胞体外发育的影响产生的猪胚胎随卵母细胞成熟和受精的条件而变化。4）从孵化的囊胚可以建立猪细胞系并在体外长期维持，并且通过将胚泡来源的细胞转移到去核卵母细胞中获得的重构胚胎具有体外发育到胚泡阶段的能力。5）包含编码绿色荧光蛋白的基因的载体6）建立了携带绿色荧光蛋白（GFP）基因的细胞向去核卵母细胞核移植的技术。

1) Basic technoloeies for the production of transgenic pigs, such as in vitro maturation and fertilization and culture of oocytes, nuclear transfer, freezing of embryos, embryo transfer, have been established.2) mitosen-activated protein kinase (MAPK) mediates the maturation-inducing signal from the cytoplasm into nucleus and induces meiosis reinitiation in porcine oocytes.3) The effect of hyaluronic acid on the development of in vitro-produced porcine embryos varies with the condition of oocyte maturation and fertilization.4) A porcine cell line can be established from the hatched blastocyst and maintained in vitro for a long period, and that reconstructed embryos obtained by transferring the blastocyst-derived cells into enucleated oocytes have the ability to develop to the blastocyst stage in vitro.5) Vector including a gene encoding green fluorescent protein (GFP gene) was developed, and techniques for selection of the cells with GFP gene was established .6) Techniques of nuclear transfer of the cells with GFP gene into enucleated oocytes was established.