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冈比亚按蚊α-微管蛋白-1b 启动子指导神经元、睾丸和发育中的 imaginal 组织特异性表达,是一种敏感的增强子检测元件。

The Anopheles gambiae alpha-tubulin-1b promoter directs neuronal, testes and developing imaginal tissue specific expression and is a sensitive enhancer detector.

机构信息

Liverpool School of Tropical Medicine, Pembroke Place, Liverpool, UK.

出版信息

Insect Mol Biol. 2012 Feb;21(1):79-88. doi: 10.1111/j.1365-2583.2011.01112.x. Epub 2011 Oct 20.

Abstract

A knowledge gap in mosquito functional genetic analysis is the dearth of characterized regulatory regions that can target tissue specific transgene expression. To broaden the tools available, a promoter region of the Anopheles gambiaeα-tubulin1b gene has been assayed following fusion to the green fluorescent protein (GFP) reporter gene and stable transformation of An. gambiae. In eight transgenic lines, the Angtub α1b regulatory region directed a core profile of tissue specific expression in the head, chordotonal organs, ventral nerve cord and testes. This profile overlaps those seen for α2-tubulin expression in Drosophila melanogaster and Bombyx mori. In addition, widespread position dependant expression was observed in other specific tissues that were unique to each line. For example, in different lines, expression was observed in larval and adult muscles, fatbody, cuticle and midgut secretory cells. The majority of genomic transgene insertions were mapped to within 10 kb of a gene, suggesting that the Angtub α1b basal promoter is particularly sensitive to enhancers and may be suitable to form the basis of a sensitive enhancer trapping construct, in combination with a binary expression system such as Gal4-UAS.

摘要

在蚊子功能基因分析方面存在知识空白,缺乏可以针对组织特异性转基因表达的特征调控区域。为了扩大可用的工具,对冈比亚按蚊α-微管蛋白 1b 基因的启动子区域进行了检测,该基因与绿色荧光蛋白(GFP)报告基因融合,并稳定转化了冈比亚按蚊。在 8 条转基因品系中,Angtub α1b 调控区在头部、感觉器官、腹神经索和睪丸中指导了核心组织特异性表达谱。该表达谱与果蝇和家蚕的α2-微管蛋白表达谱重叠。此外,还观察到在其他特定组织中存在广泛的位置依赖表达,每个品系都具有独特性。例如,在不同的品系中,在幼虫和成虫肌肉、脂肪体、角质层和中肠分泌细胞中观察到表达。大多数基因组转基因插入都映射到一个基因的 10kb 以内,这表明 Angtub α1b 基本启动子对增强子特别敏感,并且可能适合与 Gal4-UAS 等二元表达系统结合,形成敏感增强子捕获构建体的基础。

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