金纹细蛾Hsp90基因片段的克隆及其表达模式分析
发布时间：2019-03-29 06:23
【摘要】：【目的】对金纹细蛾Hsp90基因进行生物信息学和表达模式分析,为深入研究金纹细蛾PrHsp90与生长发育和抗逆行为的关系奠定基础。【方法】利用分子克隆技术获得金纹细蛾Hsp90基因片段,并对其进行生物信息学分析。同时利用RT-qPCR技术检测Hsp90在金纹细蛾不同组织、不同生长发育阶段和高温胁迫后的表达情况。【结果】获得了金纹细蛾Hsp90基因的一条长度为942bp的片段序列,命名为PrHsp90,登录号为KP671600。序列分析显示,其对应的氨基酸序列与其他昆虫的相似性很高,与棉铃虫(ADP37710.1)、斜纹夜蛾(ADK55516.1)、烟夜蛾(ADM26742.1)等鳞翅目昆虫的同源性在98%以上,表明Hsp90基因家族具有高度保守性。实时荧光定量结果表明,PrHsp90在金纹细蛾翅中的相对表达量显著高于头、胸、腹和足;金纹细蛾各个生长发育阶段都有PrHsp90表达,但其在成虫体内的相对表达量显著高于蛹期和幼虫期。高温胁迫下PrHsp90的表达水平随着温度的升高而升高,且经过35,38,41和44℃处理后的金纹细蛾,其体内PrHsp90相对表达量均显著提高,约为对照处理表达量的3~15倍。【结论】金纹细蛾PrHsp90的表达具有组织特异性,且在其生长发育的各个阶段和温度胁迫中均可能起重要作用。
[Abstract]:[objective] to analyze the bioinformatics and expression patterns of the Hsp90 gene of S. chrysophora. [methods] Hsp90 gene fragment was obtained by molecular cloning technique and analyzed by bioinformatics. [methods] the molecular cloning technique was used to obtain the Hsp90 gene fragment of PrHsp90. [methods] the molecular cloning technique was used to obtain the Hsp90 gene fragment, and the bioinformatics analysis of the gene fragment was carried out. At the same time, the expression of Hsp90 in different tissues, different stages of growth and development and after high temperature stress was detected by RT-qPCR. [results] A sequence of Hsp90 gene length 942bp, named PrHsp90, was obtained. Login number is KP671600. Sequence analysis showed that the amino acid sequence had high similarity with other insects. The homology of the amino acid sequence with other Lepidoptera insects such as ADP37710.1, ADK55516.1, ADM26742.1 was more than 98%, and the sequence analysis showed that the amino acid sequence was similar to other insects, such as Helicoverpa armigera (ADK55516.1), Spodoptera litura (ADK55516.1) and ADM26742.1. The results showed that the Hsp90 gene family was highly conserved. The results of real-time fluorescence quantitative analysis showed that the relative expression of PrHsp90 in the wings was significantly higher than that in the head, chest, abdomen and foot. The relative expression of PrHsp90 in adult was significantly higher than that in pupal and larval stages. Under high temperature stress, the expression level of PrHsp90 increased with the increase of temperature, and the relative expression of PrHsp90 increased significantly after 35,38,41 and 44 鈩，
本文编号：2449241
[Abstract]:[objective] to analyze the bioinformatics and expression patterns of the Hsp90 gene of S. chrysophora. [methods] Hsp90 gene fragment was obtained by molecular cloning technique and analyzed by bioinformatics. [methods] the molecular cloning technique was used to obtain the Hsp90 gene fragment of PrHsp90. [methods] the molecular cloning technique was used to obtain the Hsp90 gene fragment, and the bioinformatics analysis of the gene fragment was carried out. At the same time, the expression of Hsp90 in different tissues, different stages of growth and development and after high temperature stress was detected by RT-qPCR. [results] A sequence of Hsp90 gene length 942bp, named PrHsp90, was obtained. Login number is KP671600. Sequence analysis showed that the amino acid sequence had high similarity with other insects. The homology of the amino acid sequence with other Lepidoptera insects such as ADP37710.1, ADK55516.1, ADM26742.1 was more than 98%, and the sequence analysis showed that the amino acid sequence was similar to other insects, such as Helicoverpa armigera (ADK55516.1), Spodoptera litura (ADK55516.1) and ADM26742.1. The results showed that the Hsp90 gene family was highly conserved. The results of real-time fluorescence quantitative analysis showed that the relative expression of PrHsp90 in the wings was significantly higher than that in the head, chest, abdomen and foot. The relative expression of PrHsp90 in adult was significantly higher than that in pupal and larval stages. Under high temperature stress, the expression level of PrHsp90 increased with the increase of temperature, and the relative expression of PrHsp90 increased significantly after 35,38,41 and 44 鈩，
本文编号：2449241
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