| 几种诱导昆虫细胞RNA干扰质粒的构建和效果比较 |
| |
| 引用本文： | 唐盛高,尹隽,钟江. 几种诱导昆虫细胞RNA干扰质粒的构建和效果比较[J]. 复旦学报(自然科学版), 2006, 45(6): 795-799 |
| |
| 作者姓名： | 唐盛高 尹隽 钟江 |
| |
| 作者单位： | 复旦大学,生命科学学院,微生物学与微生物工程系,上海,200433;复旦大学,生命科学学院,微生物学与微生物工程系,上海,200433;复旦大学,生命科学学院,微生物学与微生物工程系,上海,200433 |
| |
| 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.
|
| 关 键 词： | RNA干扰 昆虫细胞 质粒 启动子 杆状病毒 |
| 文章编号： | 0427-7104(2006)06-0795-05 |
| 修稿时间： | 2006-05-17 |
Construction and Comparison of Plasmids for the Induction of RNA Interference in Insect Cells |
| |
| TANG Sheng-gao,YIN Juan,ZHONG Jiang. Construction and Comparison of Plasmids for the Induction of RNA Interference in Insect Cells[J]. Journal of Fudan University(Natural Science), 2006, 45(6): 795-799 |
| |
| Authors: | TANG Sheng-gao YIN Juan ZHONG Jiang |
| |
| 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. |
| |
| Keywords: | RNA interference insect cells plasmid promoter baculovirus |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ |
|
