中文摘要：褐飞虱和稻黑尾叶蝉是两种刺吸式害虫，已在东亚地区引起许多破坏性的农业灾害，它们可导致像“hopper burn”这样的病害，并传播植物病毒。最近已有连续培养这两种昆虫细胞系的报道。但是，其外源蛋白表达系统尚未建立。本研究确定了来自褐飞虱的13个微管蛋白基因和3个肌动蛋白基因，以及来自稻黑尾叶蝉的1个微管蛋白基因和2个肌动蛋白基因。此外，通过生物信息学方法和5'-RACE分析对这些基因假定的启动子区域进行分析，并通过促进增强型绿色荧光蛋白在三个昆虫细胞系S2、Sf9和BmN中的表达来评估启动子强度。本研究确定了来自褐飞虱和稻黑尾叶蝉的三个自身启动子并对其进行功能性特征描述，这将有助于适合这两种害虫的外源蛋白表达系统的建立。
外文摘要：The brown planthopper, Nilaparvata lugens (N. lugens, Hemiptera: Delphacidae), and the green rice leafhopper, Nephotettix cincticeps (N. cincticeps, Hemiptera: Cicadellidae), two sap-sucking feeders, have caused many destructive agricultural disasters in East Asia, as they can bring diseases like 'hopper burn' and transmit plant viruses. Recently, continuously-cultured cell lines from both insects have been reported. However, exogenous protein expression systems have not yet been established. Here, we identified thirteen tubulin genes and three actin genes from N. lugens, and one tubulin gene and two actin genes from N. cincticeps. Furthermore, putative promoter regions of these genes were analyzed by bioinformatic approaches and 5'-RACE assay, and the promoter strength was evaluated by driving the enhanced green fluorescent protein expression in three insect cell lines, S2, Sf9, and BmN. Finally, we identified three effective promoters (Nl_alpha Tub1 promoter, Nl_act3 promoter, and Nc_act1 promoter) among all candidates we screened. The Nc_act1 promoter showed the strongest activity, while the Nl_alpha Tub1 promoter only worked in S2 cells. In conclusion, we identified and functionally characterized three native promoters from N. lugens and N.cincticeps, which would facilitate the establishment of exogenous protein expression systems suitable for these two insect pests. (C) 2016 Elsevier Inc. All rights reserved.
外文关键词：Nilaparvata lugens；Nephotettix cincticeps；Tubulin；Actin；Promoter；Insect cell line
作者：Qian, Nannan；Zheng, Pengli；Wang, Yu；等
作者单位：北京大学
期刊名称：BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
期刊影响因子：2.371
出版年份：2016
出版刊次：2
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