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首页> 外文期刊>Biomolecular engineering >Development of a heat shock inducible and inheritable RNAi system in silkworm.
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Development of a heat shock inducible and inheritable RNAi system in silkworm.

机译：蚕中热休克诱导和可遗传RNAi系统的开发。

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摘要

A heat shock inducible and inheritable RNA interference (RNAi) system was developed in the silkworm (Bombyx mori). RNAi transgenic silkworms were generated by injecting silkworm eggs with a piggyBac transposon plasmid carrying RNAi sequence against target gene driven by the Drosophila heat shock protein 70 (HSP70) promoter and the helper plasmid expressing piggyBac transposase. The transgenic EGFP gene and the endogenous eclosion hormone (EH) gene were chosen respectively as the target genes. In the RNAi transgenic silkworms, heat shock at 42 degrees C significantly and specifically reduced the expression of EGFP or EH gene in silkworms according to the corresponding RNAi targeting sequence but not in silkworms with the irrelevant RNAi sequence demonstrating the efficiency and specificity of the RNAi effect. Heat shock in the pupal stage hampered pupal-adult eclosion and reduced egg fertility in EH RNAi transgenic silkworms but not in the wild type or EGFP RNAi transgenic silkworms. The establishment of this heat inducible and inheritable conditional RNA interference system in silkworms provided an approach for the first time to dissect the functions of target genes in silkworms at different stages.
机译：在蚕（Bombyx mori）中开发了一种热休克诱导和可遗传的RNA干扰（RNAi）系统。 RNAi转基因家蚕是通过向家蚕卵中注射携带带有针对果蝇热激蛋白70（HSP70）启动子驱动的靶基因的RNAi序列的piggyBac转座子质粒和表达piggyBac转座酶的辅助质粒而产生的。选择转基因EGFP基因和内源性促性腺激素（EH）基因作为靶基因。在RNAi转基因家蚕中，在42摄氏度下热激显着，并根据相应的RNAi靶向序列特异性降低了家蚕中EGFP或EH基因的表达，但在不具有无关RNAi序列的家蚕中证明了RNAi效应的有效性和特异性。在EH RNAi转基因家蚕中，the期的热休克阻碍了up的成虫消退并降低了卵的繁殖力，而在野生型或EGFP RNAi转基因家蚕中则没有。在蚕中建立这种热诱导和可遗传的条件性RNA干扰系统，首次提供了一种在不同阶段剖析蚕中靶基因功能的方法。

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