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长角血蜱嗅觉相关分子NPC2基因的克隆与初步鉴定

Cloning and Preliminary Identification of Olfactory-related NPC2 Gene of Haemaphysalis longicornis
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摘要 NPC2蛋白是一种小分子的溶酶体蛋白,具有水溶性。在脊椎动物中,NPC2蛋白与胆固醇和脂质的结合和运输有关;在节肢动物中参与多种信息化学物质的结合和识别。为了进一步探究蜱体内NPC2蛋白的分子特性,本研究以长角血蜱为研究对象,对HL-NPC2基因进行了克隆和序列分析,构建了原核表达载体,表达后制备出鼠源多克隆抗体,同时利用实时荧光定量qPCR技术对NPC2在蜱的不同生长阶段及不同组织进行分析,后续做了嗅觉相关性试验。结果显示:HL-NPC2蛋白的ORF为468 bp,编码155个氨基酸;NPC2蛋白在长角血蜱体内以多聚体形式存在,在长角血蜱各个生长阶段和不同吸血阶段的各个组织中都有表达;嗅觉相关性试验表明NPC2蛋白可能与长角血蜱嗅觉相关。本次研究可为该蛋白的后续研究及新的基于嗅觉的蜱控制策略的设计提供理论基础。 NPC2 protein is a small molecular lysosomal protein with water solubility.In vertebrates,NPC2 protein is related to the binding and transport of cholesterol and lipids.In arthropods,NPC2 protein is involved in the binding and recognition of a variety of information chemicals.In order to further explore the molecular characteristics of NPC2 protein in ticks,the HL-NPC2 gene of Haemaphysalis longicornis was cloned and sequenced for prokaryotic expression.The resulting NPC2 protein was used to prepare mouse polyclonal antibodies.At the same time,real-time fluorescence quantitative qPCR was used to analyze NPC2 in tick tissues at different growth stages for testing olfactory correlation.The results showed that the ORF of HL-NPC2 protein was 468 bp,encoding 155 amino acids.NPC2 protein was present as a polymer in H.longicornis and expressed in all tissues of H.longicornis at different growth stages and blood-sucking stage.Olfactory correlation test showed that NPC2 protein might be related to the sense of smell of H.longicornis.This study was helpful to future study of the protein and the design of a new tick control strategy.
作者 宋亚萍 旷策嫣 曹杰 周勇志 张厚双 武军元 周金林 SONG Yaping;KUANG Ceyan;CAO Jie;ZHOU Yongzhi;ZHANG Houshuang;WU Junyuan;ZHOU Jinlin(College of Animal Science,Tarim University,Alar 843300,China;Shanghai Veterinary Research Institute,CAAS,Shanghai 200241,China)
出处 《中国动物传染病学报》 CAS 北大核心 2024年第4期153-160,共8页 Chinese Journal of Animal Infectious Diseases
基金 国家自然科学基金资助项目(31960705,32160841) 塔里木大学研究生实践创新项目(TDGRI202132)。
关键词 长角血蜱 NPC2 克隆 鉴定 Haemaphysalis longicornis NPC2 cloning identification
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