用点杂交法检测人促红细胞生成素(hEPO)的整合

Identification of hEPO gene integration into chicken genome by dot blot hybridization

目的为制作生产人促红细胞生成素的转基因鸡,把人促红细胞生成素(hEPO)基因以质粒介导用脂质体法导入鸡胚内,然后检测人促红细胞生成素基因的整合。方法以PCR方法进行筛选后,再用点杂交法进行鉴定。即把人促红细胞生成素基因PCR扩增阳性产物用地高辛标记,作为探针和鸡整个基因组进行点杂交。结果点杂交阳性结果有4个。结论得到了整合人促红细胞生成素基因的鸡胎。

Objective： To obtain transgenic chicken that can produce hEPO. Methods： The plasmid expression vector pOV-B of human erythropoietin（ hEPO） was constructed and was embedded in liposome. Then human erythropoietin （hEPO） was integrated into chicken embryos. A pair of specific primer of hEPO gene were designed to detect all samples preserved by PCR. In order to further identify whetherexogenous gene had integrated into chicken genome, dot blot hybridization was made in the positives between 4th day and 16th day, Results：There are 4 positives in the dot blot hybridization. Conclusion： Early transgenic embryos is obtained in the studv.