摘要： 克隆药材甲热激蛋白90（Heat shock protein 90，Hsp90）基因并探讨其对高温胁迫适应性的分子机制，采用生物测定法研究了药材甲的热激存活率，并采用RT-PCR技术克隆了药材甲Hsp90基因的cDNA全长序列（GenBank登录号：KX356691），命名为SpHsp90，通过实时定量PCR技术检测其在高温胁迫后的表达量变化。结果显示，药材甲成虫的存活率随温度的升高而明显下降，44℃热激处理可导致成虫全部死亡。药材甲SpHsp90基因开放阅读框全长2 172 bp，编码723个氨基酸，理论分子量和等电点分别为82.6 kDa和5.01。SpHsp90具有Hsp90蛋白家族的5个签名序列和胞质特征序列MEEVD，并与其他昆虫HSP90具有较高的氨基酸相似性。实时定量PCR结果表明，38，40，42℃高温胁迫2 h后，药材甲成虫体内SpHsp90的表达量明显高于对照组。这说明SpHsp90在药材甲抵抗高温胁迫过程中起着重要的作用。

关键词: 药材甲, 热激蛋白, 基因克隆, 表达模式

Abstract: Heat shock protein 90(Hsp90) gene was cloned and used to reveal the molecular adaption mechanisms of high temperature stress in Stegobium paniceum (Linnaeus). In this study,we tested the survival rate of S. paniceum adults to high temperatures by bioassay method. A full-length cDNA encoding Hsp90 was cloned and characterized by RT-PCR technique. This cDNA was termed as SpHsp90 (GenBank accession No. KX356691),and its expression levels under different high temperatures were detected by using quantitative Real-time PCR(qPCR). The results showed that the survival rate of S. paniceum was significantly decreased as temperature was elevated,and the adults could not survive at 44℃. The open reading frame of SpHsp90 is 2 172 bp and encoding 723 amino acid residues,with a theoretical molecular weight of 82.6 kDa and isoelectric point of 5.01. SpHSP90 had five signature sequences of HSP90 family and a C-terminal cytoplasmic character sequence(MEEVD). The deduced amino acid residues of SpHsp90 had high similarity with Hsp90 proteins from other insect species. The qPCR analysis showed that the expression level of SpHsp90 was significantly increased 2 hours after exposed to 38,40,42℃,compared to the control. The results suggest that the SpHsp90 gene may play an important role in response to high temperature stress in S. paniceum.

Key words: Stegobium paniceum, Heat shock protein, Gene cloning, Expression pattern

中图分类号: 

• Q965

• S435.67

• Q78