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马铃薯甲虫热激蛋白基因Ld-hsp90的克隆及温度胁迫下的表达

, PP. 399-406

Keywords: 马铃薯甲虫,热激蛋白90,基因克隆,生物信息学分析,实时荧光定量PCR

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Abstract:

马铃薯甲虫Leptinotarsadecemlineata是一种抗逆性极强的世界性检疫害虫,对温度胁迫具有很强的适应性,为进一步明确其对温度胁迫适应性的分子生态机制,采用RT-PCR及RACE技术克隆得到了1个hsp90基因的cDNA全长序列,命名为Ld-hsp90,并利用实时荧光定量PCR技术检测其在温度胁迫后相对表达量的变化。Ld-hsp90全长为2489bp,开放阅读框(ORF)长度为2160bp,编码719个氨基酸,理论等电点为4.98,分子量约为82.09kD,5'端与3'端非编码区长度分别为113bp和216bp。氨基酸序列中含有HSP90家族的5个签名序列及胞质特征序列MEEVD。实时荧光定量PCR检测结果显示:38℃和44℃高温热激1h,马铃薯甲虫成虫体内的Ld-hsp90的表达量均显著高于对照组,而0℃与-10℃低温处理1h后的表达量与对照组均无显著差异。研究表明Ld-hsp90的上调表达在马铃薯甲虫抗高温中起着重要的作用。

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