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Identification and characteristic analysis of the heat shock protein 70 of Hypena tristalis (Lepidoptera : Noctuidae)

机译:三角波Hypena(鳞翅目:夜蛾科)的热激蛋白70的鉴定与特征分析

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摘要

The experiment began in June 2011, and completed in December 2012 in the insects laboratory of the North-east Agricultural University, Harbin, China. The Hypena tristalis (Htri) was collected in the North-east Agricultural University Xiang Fang farm.For the purposes of research on the expression of heat shock protein 70 (HSP 70) gene of H. tristalis treated by heat stress, with changes in expression level in different induction times, RT-PCR technology was adopted for cloning to obtain a 1905 bp HSP70 fragment of H. tristalis (GenBank accession number JQ316541), 635 amino acid residues were encoded, the predicted molecular weight was about 69.6 kD, and the isoelectric point was 5.36. The online bioinformatics prediction and analysis showed that the homology of the nucleotide sequence of H. tristalis HSP 70 with that of Helicoverpa armigera (accession number HM593518) was the highest one, which was 96%; the similarity was more than 70% while comparing with other insects. A 69.6 kD heterologous protein was detected in the HSP 70 prokaryotic expression (pET/Hfn), and the protein was branded by Western blot detection. These results provide basic data for further research on the function of HSP 70 protein and for the pest control by thermal stress.
机译:该实验于2011年6月开始,并于2012年12月在中国东北哈尔滨东北农业大学昆虫实验室完成。猪三叠纪Hypena tristalis(Htri)收集于东北农业大学湘坊农场,目的是研究热应激处理的猪三叠纪热休克蛋白70(HSP 70)基因的表达及其表达变化。在不同诱导时间水平,采用RT-PCR技术进行克隆,获得了1905 bp的H. tristalis HSP70片段(GenBank登录号JQ316541),编码635个氨基酸残基,预测分子量约为69.6 kD,等电点为5.36。在线生物信息学预测和分析结果表明,三叠纪嗜热菌HSP 70与棉铃虫(登录号HM593518)的核苷酸序列同源性最高,为96%;与其他昆虫相比,相似度超过70%。在HSP 70原核表达(pET / Hfn)中检测到69.6 kD异源蛋白,并通过Western blot检测对该蛋白进行标记。这些结果为进一步研究HSP 70蛋白的功能以及通过热胁迫控制害虫提供了基础数据。

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