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对杆状病毒感染表现出不同敏感性并表达中肠基因的小菜蛾细胞系。

Cell lines from diamondback moth exhibiting differential susceptibility to baculovirus infection and expressing midgut genes.

机构信息

State Key Laboratory of Ecological Pest Control for Fujian and Taiwan Crops, Fujian Agriculture and Forestry University, Fuzhou, China.

Fujian-Taiwan Joint Centre for Ecological Control of Crop Pests, Fujian Agriculture and Forestry University, Fuzhou, China.

出版信息

Insect Sci. 2019 Apr;26(2):251-262. doi: 10.1111/1744-7917.12533. Epub 2017 Nov 8.

Abstract

Six new cell lines were established from embryonic tissues of the diamondback moth, Plutella xylostella (L.). The cell lines showed differential characteristics, including growth in attachment or in suspension, susceptibility to a baculovirus infection and expression of genes involved in the glucosinolate detoxification pathway in P. xylostella larvae. Five of the cell lines grew attached to the culture flask and one cell line grew unattached as a suspension cell line. The cell lines had population doubling times ranging from 18 to 23 h. Among five of the P. xylostella cell lines examined for infection of a nucleopolyhedrovirus from Autographa californica, AcMNPV, four cell lines were highly susceptible to AcMNPV infection, but one was only semi-permissive to AcMNPV infection. The production of two recombinant proteins, a β-galactosidase of bacterial origin and a secreted alkaline phosphatase of eukaryotic origin, in the P. xylostella cell lines was examined in comparison with that in the cell line Sf9 which is commonly used for recombinant protein production. In the P. xylostella cell lines, expression of three important midgut genes involved in the glucosinolate detoxification pathway, including the glucosinolate sulfatase genes GSS1 and GSS2 and the sulfatase modifying factor gene SUMF1, was detected. The P. xylostella cell lines developed in this study could be useful in in vitro research systems for studying insec-virus interactions and complex molecular mechanisms in glucosinolate detoxification and insect-plant interactions.

摘要

从小菜蛾(Plutella xylostella(L.))的胚胎组织中建立了 6 个新的细胞系。这些细胞系表现出不同的特征,包括附着或悬浮生长、对杆状病毒感染的敏感性以及小菜蛾幼虫中参与硫代葡萄糖苷解毒途径的基因的表达。其中 5 个细胞系附着在培养瓶上生长,1 个细胞系作为悬浮细胞系生长。这些细胞系的群体倍增时间从 18 到 23 小时不等。在所检查的 5 个小菜蛾细胞系中,有 4 个对来自加利福尼亚棉铃虫的核多角体病毒(AcMNPV)非常敏感,但有 1 个仅对 AcMNPV 感染半允许。与常用于重组蛋白生产的 Sf9 细胞系相比,研究了在小菜蛾细胞系中生产两种重组蛋白,一种是细菌来源的β-半乳糖苷酶,一种是真核来源的分泌型碱性磷酸酶。在小菜蛾细胞系中,检测到了参与硫代葡萄糖苷解毒途径的三个重要中肠基因的表达,包括硫代葡萄糖苷硫酸酯酶基因 GSS1 和 GSS2 以及硫酸酯酶修饰因子基因 SUMF1。本研究中建立的小菜蛾细胞系可用于体外研究系统,研究昆虫-病毒相互作用以及硫代葡萄糖苷解毒和昆虫-植物相互作用中的复杂分子机制。

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