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美洲棉铃象成虫致死巨大幼虫基因的分子特征及 RNA 干扰反应。

Molecular characterization and RNA interference responses of the lethal giant larvae gene in Diabrotica virgifera virgifera adults.

机构信息

Department of Entomology, Kansas State University, Manhattan, Kansas, USA.

Institute of Applied Biology, Shanxi University, Taiyuan, Shanxi, China.

出版信息

Arch Insect Biochem Physiol. 2021 Jun;107(2):e21787. doi: 10.1002/arch.21787. Epub 2021 Apr 19.

Abstract

High specificity for silencing target genes and single-copy target genes that yield clear phenotypes are two important factors for the success of RNA interference (RNAi). The lethal giant larvae (Lgl) gene appears to be an ideal gene for RNAi because RNAi can effectively suppress its expression and results in molting defects and mortality in Tribolium castaneum. To investigate the suitability of this gene for RNAi in other insects, we identified and characterized DvLgl from the western corn rootworm, Diabrotica virgifera virgifera, a species exhibiting high RNAi efficiency. DvLgl was expressed in all developmental stages and tissues investigated. The deduced DvLgl protein showed high amino-acid sequence identities and similar domain architecture to Lgls from other insect species. Despite many similarities among insect Lgls, RNAi-mediated suppression of DvLgl failed to produce a phenotype in D. v. virgifera adults. The difference in developing phenotypes could be attributed greatly to the level of gene suppression and the insect developmental stages for RNAi. These results highlight the variability in RNAi response among insects and showcase the importance of screening multiple target genes when conducting RNAi studies. Our findings are expected to help the design of future RNAi studies and future investigations of Lgl in insects.

摘要

高特异性沉默靶基因和单拷贝靶基因,从而产生清晰的表型,是 RNA 干扰 (RNAi) 成功的两个重要因素。致死性巨大幼虫 (Lgl) 基因似乎是 RNAi 的理想基因,因为 RNAi 可以有效地抑制其表达,导致赤拟谷盗 (Tribolium castaneum) 蜕皮缺陷和死亡。为了研究该基因在其他昆虫中进行 RNAi 的适宜性,我们从西方玉米根虫(Diabrotica virgifera virgifera)中鉴定并表征了 DvLgl,该物种具有很高的 RNAi 效率。DvLgl 在所有研究的发育阶段和组织中均有表达。推导的 DvLgl 蛋白与其他昆虫物种的 Lgls 具有高度的氨基酸序列同一性和相似的结构域架构。尽管昆虫 Lgls 之间有许多相似之处,但 DvLgl 的 RNAi 介导抑制未能在 D. v. virgifera 成虫中产生表型。发育表型的差异很大程度上归因于基因抑制的水平和进行 RNAi 的昆虫发育阶段。这些结果突出了昆虫中 RNAi 反应的可变性,并强调了在进行 RNAi 研究时筛选多个靶基因的重要性。我们的研究结果有望帮助设计未来的 RNAi 研究以及未来对昆虫中 Lgl 的研究。

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