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ChtVis-Tomato,一种用于在果蝇体内可视化几丁质沉积的基因报告物。

ChtVis-Tomato, a genetic reporter for in vivo visualization of chitin deposition in Drosophila.

作者信息

Sobala Lukasz F, Wang Ying, Adler Paul N

机构信息

Biology Department and Cell Biology Department, University of Virginia, Charlottesville, VA 22904, USA.

Biology Department and Cell Biology Department, University of Virginia, Charlottesville, VA 22904, USA

出版信息

Development. 2015 Nov 15;142(22):3974-81. doi: 10.1242/dev.126987. Epub 2015 Sep 22.

Abstract

Chitin is a polymer of N-acetylglucosamine that is abundant and widely found in the biological world. It is an important constituent of the cuticular exoskeleton that plays a key role in the insect life cycle. To date, the study of chitin deposition during cuticle formation has been limited by the lack of a method to detect it in living organisms. To overcome this limitation, we have developed ChtVis-Tomato, an in vivo reporter for chitin in Drosophila. ChtVis-Tomato encodes a fusion protein that contains an apical secretion signal, a chitin-binding domain (CBD), a fluorescent protein and a cleavage site to release it from the plasma membrane. The chitin reporter allowed us to study chitin deposition in time lapse experiments and by using it we have identified unexpected deposits of chitin fibers in Drosophila pupae. ChtVis-Tomato should facilitate future studies on chitin in Drosophila and other insects.

摘要

几丁质是N-乙酰葡糖胺的聚合物,在生物界中含量丰富且广泛存在。它是表皮外骨骼的重要组成部分,在昆虫生命周期中起关键作用。迄今为止,由于缺乏在活生物体中检测几丁质的方法,角质层形成过程中几丁质沉积的研究受到了限制。为了克服这一限制,我们开发了ChtVis-Tomato,一种果蝇体内几丁质报告基因。ChtVis-Tomato编码一种融合蛋白,该蛋白包含一个顶端分泌信号、一个几丁质结合结构域(CBD)、一种荧光蛋白和一个从质膜释放它的切割位点。几丁质报告基因使我们能够在延时实验中研究几丁质沉积,通过使用它,我们在果蝇蛹中发现了几丁质纤维的意外沉积。ChtVis-Tomato应该会促进未来对果蝇和其他昆虫中几丁质的研究。

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