人红细胞生成素乳腺特异性表达载体构建及其可行性检验方法的建立

CONSTRUCTION OF MAMMARY-SPECIFIC VECTOR OF HUMAN ERYTHROPOIETIN AND ESTABLISH OF ITS FEASIBILITY DETECTION METHOD

将大鼠乳清酸蛋白（ＷＡＰ）基因的上游调控序列与人红细胞生成素基因组ＤＮＡ融合，构建了ｐＷＡＰ－ＥＰＯ－ＷＡＰ３乳腺特异性表达载体，该载体经脂质体包裹后，通过显微外科技术，经乳腺导管注入妊娠中后期大鼠乳腺中进行表达。经ＥＬＩＳＡ检测，大鼠乳汁中有ＥＰＯ表达，外周血网织红细胞计数法测定表达的ＥＰＯ具有体内生物活性。

ammary-specific expression vector of pWAP-EPO-WAP3was constructed with WAP gene promoter and erythropoietin genomic DNA. By techinique of micro-surgery, the vector mixed with lyposome was injected into mammary gland of mid-term and later stage of pregnant rats through mammary gland center duct. By ELISA detection, there was expression of EPO in rats milk. The measurement of reticulocyte numbers showed the expressive EPO in milk have biological activity in vivo. This showed that mammary gland duct injection was an ideal way that assessed the feasibility of mammary-specific expression vector.