【摘要】 目的 :构建受胰岛素、葡萄糖双向调节的胰岛素分泌调控基因的载体。方法 :1 用酚氯仿抽提法提取SD大鼠肝脏基因组DNA ,用P1、P2为引物 ,PCR扩增胰岛素生长因子结合蛋白 - 1启动子 (IGFBP - 1) ;2 选用pUC118作为克隆载体 ,用分子克隆技术亚克隆三倍体葡萄糖反应元件 (GLRE) 3;3 运用基因重组技术将IGFBP - 1启动子定向插入 (GLRE) 3的下游。结果 :1 PCR扩增出 4 2 0bp的目的DNA片断 ,与文献IGFBP - 1DNA大小一致 ;2 重组体序列分析显示 ,克隆的基因序列和文献报道的(GLRE) 3及IGFBP - 1基因序列一致。结论 :成功构建携胰岛素调控基因系统的载体pUC118(GLRE) 3-IGFBP - 1。为 1型糖尿基因治疗的进一步实验研究奠定了基础更多还原

【Abstract】 Objective:To construct an insulin gene regulated system,which can been stimulated by glucose and inhibited by insulin,in order to lay groundwork for constructing regulated insulin expression vector.Methods:The inhibitory insulin response element of the insulin-like growth factor binding protein-1 (IGFBP-1) basal promoter was amplified by PCR,Choosing pUC118 as the clone vector,we constructed three copies of GLRE by sub-clone technology,and then inserted (GLRE)3 directly upstream of IGFBP-1 by recombinant technology.Results:The DNA sequences of the recombinant gene(pUC118(GLRE)3-IGFBP-1) was the same as reported previously.Conclusion:Construction of recombinant vector caring conditioner for human proinsulin gene are obtained.更多还原