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:

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。

Abstract:

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.