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首页> 外文期刊>Weed Biology and Management >Molecular cloning, characterization, and heterologous expression analysis of heat shock protein genes (hsp70 and hsp90) of the invasive alien weed, Ageratina adenophora (Asteraceae)
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Molecular cloning, characterization, and heterologous expression analysis of heat shock protein genes (hsp70 and hsp90) of the invasive alien weed, Ageratina adenophora (Asteraceae)

机译：入侵外来杂草紫茎泽兰（菊科）热休克蛋白基因（hsp70和hsp90）的分子克隆，表征和异源表达分析

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摘要

An invasive alien weed, Ageratina adenophora, has become widespread in the southwest of China due to its high environmental adaptation. In the present study, in order to understand the molecular aspects of resistance to heat and cold stress, we investigated the characterization and expression of heat shock protein genes (designated as Aahsp70 and Aahsp90) in A. adenophora. The cDNA of A. adenophora Aahsp70 and Aahsp90 was cloned by the combination of homology cloning and rapid amplification of cDNA ends (RACE). The Basic Local Alignment Search Tool analyses revealed that Aahsp70 and Aahsp90 shared high levels of identity with corresponding proteins from other species. Hybridization was employed to examine the expression patterns of Aahsp70 and Aahsp90. The relative expression levels of the Aahsp70 and Aahsp90 transcripts were both up-regulated and reached maximal levels at 2-4 h after heat shock. Then, Aahsp70 dropped progressively to the original level at similar to 4 h after heat shock. The transcriptional changes of Aahsp70 and Aahsp90 both were more obvious under cold stress. Recombinant Aahsp70 and Aahsp90 improved the viability, in comparison with the control cultures, of Escherichia coli under stress conditions. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of the cell lysates suggested that the protective effect in vivo was related to increased thermo-stability of the soluble cytosolic proteins. This study emphasizes that the relationship between, and regulation of, heat shock proteins might incorporate variable strengths to increase the adaptation of A. adenophora.
机译：一种入侵性外来杂草，紫茎泽兰，由于其高度的环境适应性而在中国西南地区广泛传播。在本研究中，为了了解抗热和冷胁迫的分子方面，我们调查了拟南芥中热休克蛋白基因（分别称为Aahsp70和Aahsp90）的表征和表达。通过同源克隆和cDNA末端快速扩增（RACE）的结合，克隆了紫茎泽兰Aahsp70和Aahsp90的cDNA。基本局部比对搜索工具的分析表明，Aahsp70和Aahsp90与其他物种的相应蛋白质具有高度同一性。杂交被用来检查Aahsp70和Aahsp90的表达模式。热休克后2-4小时，Aahsp70和Aahsp90转录物的相对表达水平均被上调并达到最大水平。然后，Aahsp70在热冲击后约4小时逐渐下降到原始水平。在冷胁迫下，Aahsp70和Aahsp90的转录变化均更为明显。与对照培养相比，重组Aahsp70和Aahsp90在压力条件下提高了大肠杆菌的生存能力。细胞裂解物的十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分析表明，体内的保护作用与可溶性胞质蛋白的热稳定性提高有关。这项研究强调，热休克蛋白之间的关系和调控可能结合可变的强度，以增加紫茎泽兰的适应性。

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