【摘 要】
：
本研究的目的是通过转基因体细胞克隆的方法,将山羊生长激素在乳腺中特异性表达,生产转生长激素奶羊,改善乳腺的发育,促进奶产量.首先通过PCR的方法,根据Genebank上已发表的序列(GENEBANK登陆号:Z33881,Y00767),设计了三对引物,利用高保真酶克隆得到β-乳球蛋白(β-LG)5’端2284bp的片段、β-LG3’端1794bp片段和山羊生长激素CDS序列654bp片段；经序列分
【机 构】
：
南京农业大学动物医学院,南京210095 上海转基因研究中心,上海201210
【出 处】
：
中国畜牧兽医学会动物解剖学及组织胚胎学分会第十七次学术研讨会
论文部分内容阅读
本研究的目的是通过转基因体细胞克隆的方法,将山羊生长激素在乳腺中特异性表达,生产转生长激素奶羊,改善乳腺的发育,促进奶产量.首先通过PCR的方法,根据Genebank上已发表的序列(GENEBANK登陆号:Z33881,Y00767),设计了三对引物,利用高保真酶克隆得到β-乳球蛋白(β-LG)5’端2284bp的片段、β-LG3’端1794bp片段和山羊生长激素CDS序列654bp片段；经序列分析表明β-LG 5’端和3’端序列与已公布的β -LG基因序列具有同源性99％；然后以真核增强型绿色荧光表达载体pIRES-EGFP为骨架,将克隆得到β-LG5端、生长激素和β-LG3端序列,连接到真核表达载体pIRES2-EGFP上,经菌落PCR及酶切验证其插入方向正确,序列测定构建载体上的序列无碱基突变,命名为pcGH-20.
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