家蚕胚胎发育时期的RNA干涉研究

Study on The RNA Interference of Bombyx mori Embryo

通过导入特定基因的dsRNA特异性地关闭该基因的功能 ,由此产生的现象为RNA干涉 .为在家蚕胚胎发育时期建立有效的RNAi技术体系 ,在前人的基础上以家蚕第三白卵基因Bmwh3为材料 ,建立了有效的RNAi技术体系 ,结果表明 ,成功诱导第三白卵突变表型的有效注射时间为产卵后 8h内 ,wh3dsRNA的有效浓度须大于 2 0 g/L .在发育胚胎的第三天注射wh3dsRNA ,同样可诱导第三白卵突变的另一表型———半透明蚁蚕 ,根据实验结果初步推测 ,Bmwh3不仅参与眼色素和卵浆液膜色素前体的转运 ,还可能参与胚胎体壁色素前体的转运 .

Double-stranded RNA (dsRNA) can induce sequence-specific inhibition of gene function, this phenomenon is termed RNA interference (RNAi). An effective RNAi method of silkworm (Bombyx mori) embryo by introduction of double-stranded RNA corresponding to the silkworm Bmwh3 gene into the preblastoderm eggs of wild-type has been established based on the previous research. The results showed that the RNAi was successful when injected Bmwh3 dsRNA into the preblastoderm eggs within 8h after laid of wide-type silkworm and the concentration of the Bmwh3 dsRNA must be high to 2.0 g/L. It can also induce the other 0 mutant phenotype translucent larvae when injected Bmwh3dsRNA into the third day of developed eggs. A conclusion can be primarily drawn that the Bmwh3 is not only involved in the transport of the precursors of the ommochrome and pteridine pigments to the eye and egg, but also involved in the transport of the precursors of the pigments to the larvae skin.