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昆虫烟夜蛾几丁质合成酶 B 的基因、表达模式和亚细胞定位。

The gene, expression pattern and subcellular localization of chitin synthase B from the insect Ostrinia furnacalis.

机构信息

Department of Bioscience and Biotechnology, Dalian University of Technology, Dalian 116024, China.

出版信息

Biochem Biophys Res Commun. 2011 Jan 7;404(1):302-7. doi: 10.1016/j.bbrc.2010.11.111. Epub 2010 Dec 8.

Abstract

Insect midgut peritrophic membrane (PM) is a functional structure that protects insects against chemical damage and microorganism infection. The essential component in PM is chitin and its synthesis is catalyzed by Class B chitin synthase (CHSB), which plays a unique role in chitin-containing organisms and thus represents a potential target for eco-friendly pesticides. cDNA and gDNA of CHSB from a widely spread pest Ostrinia furnacalis (OfCHSB) were obtained and their sequences and transcription patterns were characterized. Results indicated that OfCHSB may be indirectly stimulated by ecdysone because the binding sites of only early ecdysone-inducible elements (BR-C and E74A) rather than ecdysone response elements (EcR and USP) were found within the core promoter of OfCHSB. In addition, the transcripts of OfCHSB increased in vivo at the feeding stage of the 4th and 5th instar larvae. The subcellular localization of OfCHSB was studied using an insect midgut cell line. Puncta structures of the recombinant OfCHSB were observed co-localized with Golgi marker Man II-GFP, suggesting a possible localization of chitin synthases under physiological conditions.

摘要

昆虫中肠围食膜(PM)是一种功能性结构,可保护昆虫免受化学损伤和微生物感染。PM 的主要成分是几丁质,其合成由 B 类几丁质合酶(CHSB)催化,CHSB 在含几丁质的生物中发挥独特作用,因此代表了环保型杀虫剂的潜在靶标。获得了广泛分布的害虫烟夜蛾(OfCHSB)的 CHSB 的 cDNA 和 gDNA,并对其序列和转录模式进行了表征。结果表明,OfCHSB 可能受到蜕皮激素的间接刺激,因为在 OfCHSB 的核心启动子中仅发现了早期蜕皮激素诱导元件(BR-C 和 E74A)的结合位点,而不是蜕皮激素反应元件(EcR 和 USP)。此外,在第 4 和第 5 龄幼虫的取食阶段,体内 OfCHSB 的转录物增加。使用昆虫中肠细胞系研究了 OfCHSB 的亚细胞定位。观察到重组 OfCHSB 的斑点结构与高尔基标记 Man II-GFP 共定位,表明在生理条件下几丁质合酶可能存在定位。

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