【摘要】 构建了四种可以在昆虫细胞中表达形成dsRNA的质粒,比较了其诱导RNA干扰,抑制目标基因———绿色荧光蛋白(GFP)基因表达的效果.四种质粒都不同程度地抑制了质粒介导的GFP表达,其中尤以单个果蝇hsp70启动子表达反向重复序列,并带有杆状病毒AcMNPV增强子hr5的质粒效果最佳,使GFP表达量下降到原来的3.9%.当用于抑制由杆状病毒介导的GFP表达时,以两个相向排列的AcMNPV ie1基因启动子的构造有明显的抑制效果.这些结果对于在昆虫细胞中有效地利用RNA i研究基因功能有参考意义.更多还原

【Abstract】 Four plasmids(pDHP-gfp,pDie1p-gfp,pHP-Dgfp,and pHPhr5-Dgfp) were constructed to express dsRNA in insect cells,and their ability to induce cellular RNA interference against the expression of green fluorescence protein(GFP) were compared.All four constructs inhibited reporter plasmid-mediated GFP expression to some degree,with the best construct to be the one with a single drosophila hsp70 promoter leading the expression of a long palindrome sequence of GFP,together with an AcMNPV enhancer hr5.GFP expression was found to be 3.9% of the control.The construct with two opposing baculovirus ie1 promoters was found to inhibit baculovirus-mediated GFP expression.These results may be helpful for the efficient application of RNAi technology in insect science.更多还原