Tisll is a Drosophila homologue of mammalian TTP which regulates the expressions of many key genes of mammals at the post-transcriptional level. But there is no available knockout or knockdown Drosophila suitable for studying Tisll function. To obtain high efficiency Tis11 gene interference Drosophila contaning Tis 11 mRNA interference sequence driven by actin or heat shock protein promoter, the GAL4 Drosophila strains, whose GAL4 expressions are driven by actin or heat shock promoter, were hybridized with the UAS Drosophila strains containing Tis 1l mRNA interference sequence. The Drosophila which carry both the GAM gene and UAS sequence were collected from Fl based on phenotype. Total RNA was extracted, and then reversely transcribed. Using specifically designed Tisll primers, the Tis1l mRNA expression were measured by real-time PCR. Compared with the control group which can not express Tis11 gene interference sequence, the Fl contaning 7M1 mRNA interference sequence showed significantly decreased levels of Tis 11 mRNA in vivo. The Fl contaning Tis 11 mRNA interference sequence showed remarkable interference effect on Tis 11 mRNA expression. Tis 11 mRNA interference Drosophila were successfully obtained.%TTP在哺乳动物许多关键基因表达的转录后水平上起调控作用,Tis11是TTP蛋白在果蝇中的同源物.目前还没有现成的可用于研究Tis11功能的基因敲除或敲低的果蝇.为了获得肌动蛋白启动子或者热激蛋白启动子驱动表达Tis11 mRNA干扰序列的具有较高干扰效率的Tis11基因干扰果蝇,将肌动蛋白启动子或者热激启动子驱动表达的GALA果蝇品系与融合有Tis11 mRNA干扰序列的UAS品系杂交,收集同时带有GALA基因和UAS序列的子一代果蝇.提取所收集果蝇的总RNA,将其中的mRNA逆转录成cDNA,并设计检测Tis11基因的特异性引物,然后通过Real-time PCR检测Tis11 mRNA的表达情况.结果显示所收集的能表达Tis11基因干扰序列的子一代果蝇与不能表达Tis11基因干扰序列的对照果蝇相比,其体内Tis11 mRNA的表达水平下降明显.收集的果蝇其体内所表达的干扰序列对Tis11 mRNA干扰效果显著,我们成功获得了Tis11基因的RNA干扰果蝇.
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