Characterization of an iron-inducible Haemaphysalis longicornis tick-derived promoter in an Ixodes scapularis-derived tick cell line (ISE6)
Characterization of an iron-inducible Haemaphysalis longicornis tick-derived promoter in an Ixodes scapularis-derived tick cell line (ISE6)
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DOI：
10.1186/s13071-019-3574-9
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发表时间：
2019-06-25
影响因子：
3.2
通讯作者：
Tanaka, Tetsuya
中科院分区：
文献类型：
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作者：
Hernandez, Emmanuel Pacia;Kusakisako, Kodai;Tanaka, Tetsuya
背景蜱是致病病原体的重要载体。随着对化学杀螨剂耐药性的增强，需要采用替代方法来控制蜱虫。基因操作已成功控制蚊子和蚊媒疾病，现在被视为控制蜱和蜱传播病原体的候选方法。我们之前的研究已经鉴定了长角血蜱中的肌动蛋白和铁蛋白启动子区域。结果在这里，对来自长角血蜱的铁蛋白衍生启动子进行了计算机表征，并鉴定了核心启动子序列及其一些重要组件。创建启动子区域的几个截短部分并将其插入报告质粒中以确定其活性的重要组成部分。使用实验报告基因和对照报告基因通过双荧光素酶测定测量肩胛硬蜱细胞系 (ISE6) 上截短启动子的活性。为了诱导启动子的活性，将转染的 ISE6 细胞暴露于硫酸亚铁。当暴露于 1 mM 硫酸亚铁时，639 个核苷酸截短的启动子在 ISE6 细胞上显示出最高活性。结论在这项研究中，我们鉴定了铁诱导蜱启动子，它可能是开发控制蜱和蜱传病原体的基因操作系统的宝贵工具。
BackgroundTicks are important vectors of disease-causing pathogens. With the rise of resistance to chemical acaricides, alternative methods in tick control are warranted. Gene manipulation has been successful in controlling mosquitoes and mosquito-borne diseases and is now looked upon as a candidate method to control ticks and tick-borne pathogens. Our previous study has identified the actin and ferritin promoter regions in the Haemaphysalis longicornis tick.ResultsHere, the ferritin-derived promoter from the H. longicornis tick was characterized in silico, and the core promoter sequences and some of its important components were identified. Several truncations of the promoter region were created and inserted to a reporter plasmid to determine the important components for its activity. The activities of the truncated promoters on the Ixodes scapularis tick cell line (ISE6) were measured via a dual luciferase assay using experimental and control reporter genes. To induce the promoter's activity, transfected ISE6 cells were exposed to ferrous sulfate. The 639 nucleotides truncated promoter showed the highest activity on ISE6 cells when exposed to 1 mM ferrous sulfate.ConclusionIn this study, we characterized an iron-inducible tick promoter that could be a valuable tool in the development of a gene-manipulation system to control ticks and tick-borne pathogens.