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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
摘要点击 36  全文点击 29  投稿时间:2016-11-21  修订日期:2016-12-23
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DOI编号   
中文关键词   药材甲  热激蛋白60  基因克隆  温度胁迫  表达模式
英文关键词   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)
作者单位E-mail
朱晓晔 贵州师范大学 贵阳学院 18886074373@163.com 
丁天波 青岛农业大学  
许抗抗 贵阳学院  
严毅 贵阳学院  
李灿 贵阳学院  
杨文佳 贵阳学院 yangwenjia10@126.com 
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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