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公开(公告)号:KR1020070024069A
公开(公告)日:2007-03-02
申请号:KR1020050078585
申请日:2005-08-26
Abstract: A method for producing transgenic pig capable of expressing colony stimulating factor(CSF) gene in the mammary gland is provided to be able to easily obtain the transgenic pig, which is able to solve the problems of post-translational modification or mass-production and is used as a supplemental agent of treating leukemia and an immuno-potentiating agent, by expressing hGM-CSF in mammary gland. The method comprises the steps of: (a) isolating somatic cell from pig fetus on days 40-80 pregnancy; (b) after preparing an expression vector including a promoter specific to mammary gland such as alphas1-casein, beta-casein, beta-lactoglobulin and whey acidic protein; and a polynucleotide encoding the CSF such as hGM-CSF, introducing it into the somatic cell; (c) after selecting the expression vector-introduced clone somatic cell, culturing it; (d) removing nucleus of ovum collected from sow and fusing it with the clone somatic cell; and (e) transplanting the fused nuclear transfer embryos to surrogate mother pig and delivering piglets.
Abstract translation: 提供了能够在乳腺中表达集落刺激因子(CSF)基因的转基因猪的制备方法,以便能够容易地获得能够解决翻译后修饰或批量生产的问题的转基因猪,并且是 用作通过在乳腺中表达hGM-CSF来治疗白血病和免疫增强剂的补充剂。 该方法包括以下步骤:(a)在妊娠40-80天从胎儿分离体细胞; (b)在制备包括对乳腺特异性的启动子如α-α-酪蛋白,β-酪蛋白,β-乳球蛋白和乳清酸性蛋白质的表达载体后; 和编码CSF的多核苷酸,例如hGM-CSF,将其引入体细胞; (c)选择表达载体导入的克隆体细胞后进行培养; (d)去除从母猪收集的卵子的细胞核并与克隆体细胞融合; 和(e)将融合的核转移胚胎移植到替代母猪和递送仔猪。