大鼠肾小球系膜细胞表达人白细胞介素10

Expression of Human Interleukin 10 in Rat Glomerular Mesangial Cells by Retroviral Vector

构建表达人白细胞介素１０的逆转录病毒重组体Ｐ（ｈＩＬ－１０）ＳＮ，经ＰＡ３１７细胞包装，Ｇ４１８筛选，用重组逆转录病毒感染大鼠肾小球系膜细胞，应用聚合酶链反应（ＰＣＲ），反转录聚合酶链反应（ＲＴ－ＰＣＲ）和ＥＦＩＳＡ检测ｈＩＬ－１０基因的整合和表达．结果显示外源性ｈＩＬ－１０基因已整合到靶细胞染色体ＤＮＡ并有效地表达，高峰在第３天，为每天每个细胞１．８４３×１０－３ｎｇ。

Retroviral recombinant pLX(hIL 10)SN was constructed and introduced it into packaging cell line PA317.After PA317 was selected by G418,using recombinant retrtoviral vector pLX(hIL 10)SN to infect rat glomerular mesangial cells.After gene transfer 72 h,DNA and RNA were prepared from rat transfected mesangial cells for the polymerase chain reaction(PCR) and the reverse transcription polyverase chaim reaction(RT PCR).Exogenous of hIL 10gene has been subcloned into retrovial vector pLXSN successfully,and it has been integrated into the chromosal DNA of the transfected masangial cells.RT PCR and ELISA showed that exogenous hIL 10 gene was experssed in the transtected mesangial cells after transfection 48 h and lasting for over 28 d.The retroviral vector pLX(hIL 10)SN shall lay foundation for researching hIL 10 mechanism and its application for curing deseases.