药材甲热激蛋白基因SpHsp60的克隆及温度胁迫下的表达
cDNA cloning of heat shock protein 60 (SpHsp60) in Stegobium paniceum and its expression under temperature stress
作者：朱晓晔(贵州师范大学生命科学学院; 贵阳学院生物与环境工程学院, 贵州省山地珍稀动物与经济昆虫重点实验室)；丁天波(青岛农业大学植物医学学院)；许抗抗(贵阳学院生物与环境工程学院, 贵州省山地珍稀动物与经济昆虫重点实验室)；严毅(贵阳学院生物与环境工程学院, 贵州省山地珍稀动物与经济昆虫重点实验室)；李灿(贵阳学院生物与环境工程学院, 贵州省山地珍稀动物与经济昆虫重点实验室)；杨文佳(贵阳学院生物与环境工程学院, 贵州省山地珍稀动物与经济昆虫重点实验室)
Author：ZHU Xiao-Ye(College of Life Science, Guizhou Normal University; Guizhou Provincial Key Laboratory for Rare Animal and Economic Insects of the Mountainous Region, College of Biology and Environmental Engineering, Guiyang University)；DING Tian-Bo(College of Plant Health and Medicine, Qingdao Agricultural University)；XU Kang-Kang(Guizhou Provincial Key Laboratory for Rare Animal and Economic Insects of the Mountainous Region, College of Biology and Environmental Engineering, Guiyang University)；YAN Yi(Guizhou Provincial Key Laboratory for Rare Animal and Economic Insects of the Mountainous Region, College of Biology and Environmental Engineering, Guiyang University)；LI Can(Guizhou Provincial Key Laboratory for Rare Animal and Economic Insects of the Mountainous Region, College of Biology and Environmental Engineering, Guiyang University)；YANG Wen-Jia(Guizhou Provincial Key Laboratory for Rare Animal and Economic Insects of the Mountainous Region, College of Biology and Environmental Engineering, Guiyang University)
收稿日期：2016-11-21 年卷（期）页码：2017,54(6):1311-1316
期刊名称：四川大学学报: 自然科学版
Journal Name：Journal of Sichuan University (Natural Science Edition)
关键字：药材甲；热激蛋白60；基因克隆；温度胁迫；表达模式
Key words：Stegobium paniceum; Heat shock protein 60; Gene cloning; Temperature stress; Expression pattern
基金项目：国家自然科学基金青年基金项目（31501649），贵州省高层次创新型人才培养（黔科合人才[2016]4020）；贵州省科学技术联合基金项目（黔科合LH字[2014]7167）；贵阳学院高层次人才科研启动费（校人才2014003）
中文摘要
本研究以中药材储藏期主要害虫药材甲为材料，采用RT-PCR技术克隆了药材甲热激蛋白60（Heat shock protein 60, Hsp60）基因的cDNA全长序列，命名为SpHsp60（GenBank登录号：KX778623）。测序结果表明，SpHsp60的开放阅读框长度为1719 bp，编码572个氨基酸，理论分子量和等电点分别为60.9 kDa和5.32。SpHsp60具有Hsp60基因家族高度保守的基序，SpHsp60编码的氨基酸序列与赤拟谷盗Tribolium castaneum和马铃薯甲虫Leptinotarsa decemlineata等昆虫的Hsp60的氨基酸序列一致性为88%。通过实时定量PCR技术检测了不同温度胁迫后该基因的表达量，结果表明：-5°C和0°C 低温处理2 h，药材甲成虫体内的SpHsp60的表达量均显著高于对照组，且42°C高温处理2 h后的表达量也显著高于对照组。说明SpHSP60与药材甲应对极端温度胁迫相关。
英文摘要
The full-length cDNA sequence of a heat shock protein 60 (Hsp60) gene from Stegobium paniceum (a major insect pest in stored Chinese medicine material) was cloned by using RT-PCR technique, and termed as SpHsp60 (GenBank accession no. KX778623). Sequence analysis showed that SpHsp60 contained an open reading frame of 1719 bp encoding 572 amino acids, with a predicated molecular weight of 60.9 kDa and isoelectric point of 5.32. SpHsp60 had the highly conserved functional motifs of the Hsp60 gene family, and SpHsp60 shares 88% amino acid sequence identities with the Hsp60 proteins from Tribolium castaneum and Leptinotarsa decemlineata. The quantitative real-time PCR results indicated that the expression of SpHsp60 was significantly increased 2 hours after exposed to -5°C and 0°C, compared to the control, and this gene was also induced after exposed to 42°C. The results suggest that the SpHsp60 gene may correlate with extreme temperature-stress tolerance in S. paniceum.
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