设计并构建了针对par-3基因的发夹RNA载体,将构建好的质粒转入大肠杆菌HT115,25℃喂食Caenorhabditis elegans(C.elegans)野生型虫体,24h后观察par-3(RNA干扰)celegans的胚胎发育情况。结果显示通过喂食形成发夹结构dsRNA的细菌可以对celegans中par-3基因进行RNA干扰,干扰率可以达到60%以上。干扰后的早期胚胎发育丧失第一次卵裂的不对称性,第二次卵裂的纺锤体方向发生改变,与par-3突变体的观察结果一致,为进一步在mex-3转基因虫体中通过RNA干扰研究基因表达打下了基础。
The hairpin dsRNA expression vector targeting par-3 gene was constructed and transformed into E.coli HT115. The development of C.elegans embryo was observed after feeded bacteria which contain par-3 dsRNA plasmids for 24 h in 25 ℃. The results showed that this construct of RNAi targeting par-3 gene delivered by feeding this bacteria is efficient. The bacteria mediated RNAi of par-3 result in the loss of the asymmetry of the C. elegans early embryo and the change of the spindle orientation in the second cleavage. This construct can be applied into the further study of gene expression in transgenetic worms via bacteria mediated RNAi.