摘要
热激蛋白(heat shock protein,HSP)是生物体应对不良环境胁迫的关键蛋白,HSP90是热激蛋白家族中的重要成员。琥珀蚕(Antheraea assamensis)是一种半驯养的经济昆虫,在生长过程中常受到高低温等不利环境条件的冲击。从琥珀蚕中克隆得到AaHsp90基因的全长cDNA序列(GenBank登录号:MK1656641),该序列长2482 bp,包括126 bp的5′非编码区(5′UTR)和202 bp的3′非编码区(3′UTR),开放阅读框为2154 bp,编码717个氨基酸,预测蛋白质分子质量为824 kD,等电点为500。AaHSP90氨基酸序列含有HSP90家族的5个特征序列及C末端的胞质保守基序MEEVD,氨基酸序列同源比对结果表明其与天蚕和柞蚕的同源序列相似度分别达到981%和990%。实时荧光定量PCR结果表明,该基因mRNA在琥珀蚕卵、幼虫期各龄及5龄第4天不同组织中均有表达;相对于对照组,在高温胁迫条件下其表达量显著上调,低温刺激条件下其表达则受到抑制。研究表明AaHsp90基因在琥珀蚕对环境温度适应中起重要作用。
Heat shock proteins(HSPs)are key elements in response to environmental stress.HSP90 is the important member of HSPs.As a semi-domesticated economic insect,Antheraea assamensis is always subject to low or high temperature stress in the process of growth.Therefore,ful-length cDNA sequence of AaHsp90 gene(GenBank accession No.MK165664.1)was cloned from A.assamensis in this study.This gene is 2482 bp in length,containing 126 bp 5′-untranslated region(5′UTR)and202 bp 3′-untranslated region(3′UTR).Its open reading frame is 2154 bp encoding 717 amino acid with predicted molecular mass of 82.4 kD and isoelectric point of 5.00.Besides,amino acid sequence of AaHSP90 contains 5 signature sequences of HSP90 family and a C-terminal cytoplasmic characteristic sequence(MEEVD).Amino acid sequence alignment results indicated that HSP90 of Antheraea assamensis shared 98.1%similarity with Antheraea yamamai and 99.0%similarity with Antheraea pernyi.Quantitative real-time PCR results showed that AaHsp90 was expressed in egg and different tissues of A.assamensis in different larval stages and of the 4 th day 5 th instar larvae.Compared with the control group,expression level of AaHsp90 was significantly upregulated under high temperature stress,while suppressed under low temperature stress.The results suggested that AaHsp90 of A.assamensis plays an important role in its adaptation to high or low temperature.
作者
刘增虎
钟健
杨伟克
李琼艳
陈安利
董占鹏
Liu Zenghu;Zhong Jian;Yang Weike;Li Qiongyan;Chen Anli;Dong Zhanpeng(Institute of Sericulture and Apiculture,Yunnan Academy of Agricultural Sciences,Mengzi Yunnan 661101,China)
出处
《蚕业科学》
CAS
CSCD
北大核心
2020年第1期59-66,共8页
ACTA SERICOLOGICA SINICA
基金
国家自然科学基金项目(31360586)
云南省应用基础研究面上项目(2017FB069).
关键词
琥珀蚕
热激蛋白90
克隆
表达
实时荧光定量PCR
温度胁迫
Antheraea assamensis
Heat shock protein 90(HSP90)
Cloning
Expression
Quantitative real-time PCR
Temperature stress
