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Starvation-induced autophagy in Spodoptera frugiperda Sf9 ovarian cells
Physiological Entomology ( IF 1.6 ) Pub Date : 2019-02-23 , DOI: 10.1111/phen.12279
Ruopeng Yang 1 , Xiyu Shang 2 , Chuanming Wang 1 , Mijie Li 1 , Chengyan Jiang 1 , Haiju Lu 1 , Qiong Kong 1 , Bo Gao 1 , Wanying Yang 3 , Yang Cao 3 , Kun Xie 1, 3
Affiliation  

Autophagy controls insect development and can be targeted for pest control in agriculture. In the present study, starvation‐induced autophagy is investigated in the insect species Spodoptera frugiperda. Bioinformatics analysis and a search of the EST database (http://bioweb.ensam.inra.fr/spodobase) identifies a putative ATG8 gene of S. frugiperda. To generate a biomarker of autophagosome, the DNA sequence encoding the open reading frame of this gene is amplified and cloned into a pIEX‐4‐mCherry‐EGFP‐SfATG8 recombinant vector. Sf9 cells are then transfected with this expression vector and starved in phosphate‐buffered saline solution for 4 h to induce autophagy, which is examined by LysoTracker staining (Life Technologies, Grand Island, New York), western blotting and fluorescence microscopy. The results obtained show that starvation stimulates lipidation of SfATG8‐PE and the formation of autophagosomes, providing a foundation for further research with respect to autophagy in insects.

中文翻译:

草地贪夜蛾 Sf9 卵巢细胞饥饿诱导的自噬

自噬控制昆虫发育,可作为农业害虫防治的目标。在本研究中,在昆虫种类草地贪夜蛾中研究了饥饿诱导的自噬。生物信息学分析和对 EST 数据库 (http://bioweb.ensam.inra.fr/spodobase) 的搜索确定了 S. frugiperda 的推定 ATG8 基因。为了产生自噬体的生物标志物,编码该基因开放阅读框的 DNA 序列被扩增并克隆到 pIEX-4-mCherry-EGFP-SfATG8 重组载体中。然后用该表达载体转染 Sf9 细胞并在磷酸盐缓冲盐水溶液中饥饿 4 小时以诱导自噬,通过 LysoTracker 染色(Life Technologies,Grand Island,New York)、蛋白质印迹和荧光显微镜检查。
更新日期:2019-02-23
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