Cloning and expression analysis of six small heat shock protein genes in the common cutworm, Spodoptera litura
Cloning and expression analysis of six small heat shock protein genes in the common cutworm, Spodoptera litura
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斜纹夜蛾6个小热激蛋白基因的克隆及表达分析
DOI：
10.1016/j.jinsphys.2011.03.026
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发表时间：
2011-07-01
影响因子：
2.2
通讯作者：
Kang, Le
中科院分区：
文献类型：
--
作者：
Shen, Ying;Gu, Jun;Kang, Le
小分子热休克蛋白（sHsps）在各种应激蛋白超家族中可能在结构和功能上最为多样。为了探究昆虫sHsps的多种功能，从斜纹夜蛾中肠cDNA文库中克隆了6个sHsp cDNA，并基于保守的α-晶状体结构域构建了系统发育树。通过实时定量PCR研究了它们在不同发育阶段和组织中的表达模式，以及对热诱导和20 - 羟基蜕皮酮（20E）诱导的响应。根据序列特征和系统发育关系，这6个SIHsps被分为3个独立的组：类BmHsp20.4蛋白（SIHsp19.7、20.4、20.7、20.8）、类BmHsp26.6蛋白（SIHsp20）和类BmHsp21.4蛋白（SIHsp21.4）。所有的SIHsps在马氏管中表达量最高。4个类BmHsp20.4蛋白基因受热应激上调，并随发育表现出表达变化。SIHsp20在卵和幼虫阶段的表达水平低于蛹和成虫阶段。SIHsp21.4在所有生活阶段都保持恒定的表达水平。SIHsp20.4和SIHsp20.8的表达都被20E显著上调。这些结果表明，sHsps在斜纹夜蛾中发挥多种功能：类BmHsp20.4蛋白参与热适应和发育；SIHsp20对温度应激无响应，但可能在变态中起作用；SIHsp21.4可能与热响应或发育都没有直接关系。（C）2011爱思唯尔有限公司。保留所有权利。
Small heat shock proteins (sHsps) are probably the most diverse in structure and function among the various superfamilies of stress proteins. To explore the diverse functions of insect sHsps, six sHsp cDNAs were cloned from the midgut cDNA library of Spodoptera litura, and a phylogenetic tree was constructed based on the conserved a-crystalline domains. The expression patterns in different developmental stages and tissues, as well as in response to both thermal and 20-hydroxyecdysone (20E) induction, were studied by real-time quantitative PCR. Based on sequence characteristics and phylogenetic relationships, the six SIHsps were classified into three independent groups: BmHsp20.4 like proteins (SIHsp19.7, 20.4, 20.7, 20.8), BmHsp26.6 like protein (SIHsp20), and BmHsp21.4 like protein (SIHsp21.4). All the SIHsps showed highest expression in the Malpighian tubules. The four BmHsp20.4 like protein genes were up-regulated by thermal stress and showed expression variation with development. SIHsp20 exhibited lower expression levels in both egg and larval stages than in pupal and adult stages. SIHsp21.4 retained a constant expression level during all life stages. The expression of both SIHsp20.4 and SIHsp20.8 was significantly up-regulated by 20E. These results indicate that sHsps play diverse functions in S. litura: the BmHsp20.4 like proteins are involved in both thermal adaptation and development: SIHsp20 does not respond to temperature stress but possibly plays a role in metamorphosis: SIHsp21.4 may have no direct relationship with either thermal response or development. (C) 2011 Elsevier Ltd. All rights reserved.