黏虫热休克蛋白Hsp90基因的克隆及不同温度对其诱导反应

doi:10.16409/j.cnki.2095-039x.2017.05.007

中国生物防治学报 ›› 2017, Vol. 33 ›› Issue (5): 623-630.DOI: 10.16409/j.cnki.2095-039x.2017.05.007

黏虫热休克蛋白Hsp90基因的克隆及不同温度对其诱导反应

杨静, Nasir Ali Baloch, 樊东

东北农业大学农学院, 哈尔滨 150030

收稿日期:2017-09-18 出版日期:2017-10-08 发布日期:2017-10-11
通讯作者: 樊东
作者简介:杨静,硕士研究生,E-mail:1311762737@qq.com;樊东,教授,博士生导师,E-mail:dnfd@163.com。
基金资助:
黑龙江省自然科学基金（C201415）

cDNA Cloning and Induction of Heat Shock Protein Hsp90 from Mythimna separate

YANG Jing, NASIR Ali Baloch, FAN Dong

College of Agronomy, Northeast Agricultural University, Harbin 150030, China

Received:2017-09-18 Online:2017-10-08 Published:2017-10-11

摘要/Abstract

摘要： 热休克蛋白是昆虫体内一种很重要的抗逆蛋白，Hsp90是热休克蛋白家族中的重要成员。本试验利用高通量测序法获得Hsp90的cDNA的全长序列，通过Real-time PCR探讨黏虫各龄期、组织和黏虫3龄幼虫受到不同高、低温胁迫不同时间的Hsp90基因的相对表达量。在黏虫体内得到Hsp90的cDNA全长序列（GenBank登录号MF773751）2422 bp，命名为MsHsp90，编码717个氨基酸，分子量约为82.576 ku，等电点是4.98，序列包含有Hsp90的保守序列，与鳞翅目夜蛾科的甜菜夜蛾、苜蓿夜蛾、斜纹夜蛾亲缘关系较近，且氨基酸序列相似性都在98%以上。Real-time PCR结果显示黏虫的不同发育阶段和组织中均有MsHsp90表达，其中2龄幼虫和后肠的相对表达量最高。40℃处理6 h的相对表达量最高，处理12、24、48 h时，20℃的相对表达量最高。上述分析可知，MsHsp90相对表达量的高低表明在黏虫不同组织，不同发育阶段和不同时间、温度处理中发挥的作用存在差异。

关键词: 黏虫, 热休克蛋白Hsp90, 温度胁迫

Abstract: Heat shock proteins are key elements of the organism's adaptive response to adverse environments. Hsp90 has an important place in the Hsp superfamily. Full length of Hsp90 cDNA sequence was obtained from Mythimna separata by using high-throughput sequencing. Hsp90 mRNA transcripts in M. separata were examined at various developmental stages and in different tissues based on real-time PCR. Hsp90 mRNA expression levels were also examined to different temperature treatments. The results showed that the cDNA sequence of Hsp90 in M. separata is 2422 bp and encodes 717 amino acids. The molecular mass is about 82.576 ku, and isoelectric point is about 4.98. The amino acid sequence includes signature sequences of Hsp90. Sequence alignment showed that MsHsp90 shared more than 98% identity with Hsp90 sequences reported in Spodoptera exigua, Spodoptera litura and Heliothis viriplaca.The cDNA sequence was designated as MsHsp90 (GenBank accession No. MF773751). The relative expression of MsHsp90 in the 2nd instar larvae and hindgut was higher than that in other developmental stages and tissues. The relative expression level of MsHsp90 in M. separata varies in different temperature stress and time periods. Based on the analysis above, the relative expression level of MsHsp90 in different tissues, at different developmental stages and in different temperature induction indicated that MsHsp90 may play different roles in M. separata.

Key words: Mythimna separata, heat shock protein 90, temperature induction

中图分类号:

S433.4

杨静, Nasir Ali Baloch, 樊东. 黏虫热休克蛋白Hsp90基因的克隆及不同温度对其诱导反应[J]. 中国生物防治学报, 2017, 33(5): 623-630.

YANG Jing, NASIR Ali Baloch, FAN Dong. cDNA Cloning and Induction of Heat Shock Protein Hsp90 from Mythimna separate[J]. journal1, 2017, 33(5): 623-630.

参考文献

[1] Shen Y, Gu J, Huang L H, et al. Cloning and expression analysis of six small heat shock protein genes in the common cutworm, Spodoptera litura[J]. Journal of Insect Physiology, 2011, 57(7):908-914.
[2] Ritossa F M. A new puffing pattern induced by temperature shock and DNP in Drosophila[J]. Experiantia, 1962, 18(1):571-573.
[3] Tissieres A, Mitchell H K, Tracy U M. Protein synthesis in salivary glands of Drosophila melanogaster:relation to chromosome puffs[J]. Journal of Molecular Biology, 1974, 84(3):389-398.
[4] Marber M, Mestril R, Chis H, et al. Overexpression of the rat inducible 70 kDa heat stress protein in a transgenic mouse increases the resistance of the heart to ischemic injury[J]. Journal of Clinical Investigation, 1995, 95(4):1446-1456.
[5] Weitzel G, Pilatus U, Rensing L. Similar dose response of heat shock protein synthesis and intracellular pH change in yeast[J]. Experimental Cell Research, 1985, 159(1):252-256.
[6] Levison W, Oppermann H, Jackson J. Transitionseries metals and sulfhydryl reagents induce the synthesis of four proteins in eukaryotic cells[J]. Biochimica et Biophysica Acta, 1980, 606(1):170-180.
[7] Shu Y H, Du Y, Wang J W. Molecular characterization and expression patterns of Spodoptera litura heat shock protein 70/90, and their response to zinc stress[J]. Comparative Biochemistry and Physiology Part A:Molecular and Integrative Physiology, 2011, 158(1):102-110.
[8] Kelley P M, Schlesinger M J. The effect of amino acid analogues and heat shock on gene expression in chicken embryo fibroblasts[J]. Cell, 1978, 15(4):1277-1286.
[9] 陈亚琼, 肖调红, 周浙昆. 热激蛋白与生物环境适应及进化的关系[J]. 自然科学进展, 2006, 16(9):1066-1073.
[10] Parsell D A, Lindquists S. Heat shock proteins and stress tolerance[M]//Morimoto R I, Tissieres A, Genrgoponlous C, eds. The Biolog of Heat Shock Proteins and Molecular Chaperones. New York:Coldspring Harbor Laboratory Press, 1994, 457-494.
[11] Bale J S, Masters G J, Hodkinson I D, et al. Herbivory in global climate change research:direct effects of rising temperature on insect herbivores[J]. Global Change Biology, 2002, 8(1):1-16.
[12] 李玉婷, 赵奇, 房冰, 等. 禾谷缢管蚜热激蛋白Hsp90基因的克隆和表达分析[J]. 植物保护学报, 2017, 44(1):16-23.
[13] 张青, 陆明星, 祝树德. 灰飞虱热激蛋白Hsp90基因的克隆、序列分析与表达模式[J]. 昆虫学报, 2014, 57(7):777-786.
[14] Xu Q, Zou Q, Zheng H Z, et al. Three heat shock proteins from Spodoptera exigua:gene cloning, characterization and comparative stress response during heat and cold shocks[J]. Comparative Biochemistry and Physiology Part B:Biochemistry and Molecular Biology, 2012, 159(2):92-102.
[15] 申建茹, 李明福, 陈乃中, 等. 苹果蠹蛾热激蛋白Hsp90基因的克隆及热胁迫下的表达分析[J]. 昆虫学报, 2011, 54(11):1236-1248.
[16] 蒋健, 李祥瑞, 张云慧, 等. 马铃薯甲虫热激蛋白基因Ldhsp90的克隆及温度胁迫下的表达[J]. 植物保护学报, 2013, 40(5):399-406.
[17] Zhang Y, Zhang Z, Li C, et al. Seasonal migratory behavior of Mythimna separata (Lepidoptera:Noctuidae) in Northeast China[J]. Acta Entomologica Sinica, 2013, 56(12):1418-1429.
[18] 王冠华, 李鑫, 郭长宁, 等. 金纹细蛾Hsp90基因片段的克隆及其表达模式分析[J]. 西北农林科技大学学报(自然科学版), 2016, 44(10):114-120.
[19] 申建茹, 李明福, 陈乃中, 等. 苹果蠹蛾热激蛋白Hsp90基因的克隆及热胁迫下的表达分析[J]. 昆虫学报, 2011, 54(11):1236-1248.
[20] Zhang Y, Gu S S, Li C G, et al. Identification and characterization of novel ER-based hsp90 gene in the red flour beetle, Tribolium castaneum[J]. Cell Stress and Chaperones, 2014, 19(5):623-633.

黏虫热休克蛋白Hsp90基因的克隆及不同温度对其诱导反应

cDNA Cloning and Induction of Heat Shock Protein Hsp90 from Mythimna separate

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