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OALib Journal期刊
ISSN: 2333-9721
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朱砂叶螨乙酰胆碱酯酶基因的克隆与特征分析
DOI: 10.3724/SP.J.1145.2014.01051, PP. 751-758
Keywords: 朱砂叶螨,乙酰胆碱酯酶,基因克隆,荧光定量
Abstract:
乙酰胆碱酯酶（ache）是有机磷和氨基甲酸酯类杀虫（螨）剂的主要作用靶标，为建立以朱砂叶螨（tetranychuscinnabarinus）ache为靶点的杀螨剂体外筛选体系，本研究拟克隆朱砂叶螨ache基因，并分析其特征.采用同源基因克隆及race技术克隆基因，通过荧光定量pcr分析其表达特征.结果获得朱砂叶螨ache的cdna序列长为2512bp，其中开放阅读框（orf）为2064bp，编码687个氨基酸，n端有信号肽，剪切位点在甲硫氨酸开始的第74-75个氨基酸残基之间，c端有糖酯锚定结构，genbank登录号为agi96546.1.序列分析显示朱砂叶螨与其他蜱螨目的ache有高度的同源性，特别与同属叶螨属的二斑叶螨（tetranychusurticae）ache的同源性最高，但氨基酸序列比较分析显示两者酶的催化特性可能存在区别.朱砂叶螨不同发育期ache的转录水平比较分析显示幼螨期的表达量最高，且从幼螨期开始表达量逐渐下降，成螨期最低.本研究表明朱砂叶螨与二斑叶螨的ace1有较高的同源性，但酶特性存在差异，朱砂叶螨不同发育时期ace1基因表达水平的不同可能与不同发育期螨的习性和代谢特性有关.图7参20
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