江汉大学学报(自然科学版) ›› 2016, Vol. 44 ›› Issue (6): 490-495.doi: 10.16389/j.cnki.cn42-1737/n.2016.06.002

• 豆类植物研究 • 上一篇    下一篇

四纹豆象小分子量热激蛋白基因的鉴定与表达分析

刘琴,陈伟达,高志鹏,张雨辰,陈禅友   

  1. 江汉大学 生命科学学院;湖北省豆类(蔬菜)植物工程技术研究中心,湖北 武汉 430056
  • 出版日期:2016-12-28 发布日期:2016-12-30
  • 作者简介:刘琴(1983—),女,讲师,博士,研究方向:昆虫微生物及其生物技术。
  • 基金资助:
    湖北省豆类(蔬菜)植物工程技术研究中心开放基金项目(2016-08)

Identification and Expression Analysis of Gene of Small Heat Shock Protein in Callosobruchus maculatus

LIU Qin ,CHEN Weida,GAO Zhipeng,ZHANG Yuchen,CHEN Chanyou   

  1. School of Life Sciences,Jianghan University;Hubei Province Engineering Research Center for Legume Plants,Wuhan 430056,Hubei,China
  • Online:2016-12-28 Published:2016-12-30

摘要: 本研究利用生物信息学方法对NCBI 数据库中四纹豆象(Callosobruchus maculatus)全部热激蛋白进行分析,从中鉴定出一种中央区具有典型α-晶体蛋白质结构域的小分子量热激蛋白基因,shsp27(GenBank编号FK669241.1)。采用实时荧光定量PCR(RT-qPCR)检测了四纹豆象4龄幼虫经不同温度(4、15、30、37 ℃以及4 ℃低温处理24 h 后回复30 ℃)胁迫处理24 h 后,shsp27 mRNA 的相对表达量。分析结果表明,在4龄四纹豆象幼虫中,低温(4、15 ℃)与高温(37 ℃)胁迫处理后shsp27的相对表达量相对于对照组(30 ℃恒温培养组)均有上调响应,但对高温胁迫的响应程度略强,低温回复处理组的相对表达量也略高于对照组,但上升不显著,初步认为shsp27基因表达与四纹豆象耐寒热的适应性无直接相关性。

关键词: 四纹豆象, 小分子量热激蛋白, 温度胁迫, mRNA相对表达量, 实时荧光定量PCR

Abstract: In this paper,the author identified one particular small heat shock protein(sHSP)( shsp27 ,GenBank number FK669241.1) which contained a typical α- crystalline central domain from Callosobruchus maculatus with bioinformatics method. Then,the author used real- time quantitative PCR(RT-qPCR)to detect the mRNA relative expression levels of shsp27 in 4th larva of C. maculatus ,after being exposured to different temperature stress(4,15,30,37 ℃,and 4 ℃ 24 h then back to 30 ℃ treatments(Rec)). The results showed that the low temperature(4,15 ℃)and high temperature(37 ℃)heat stress up-regulated the relative expression of shsp27 ,and the high temperature stress upregulated it stronger,that of Rec was slightly higher than that of CK,but not significantly. It is considered that there is no correlationship between shsp27 expression and high/low temperature adaptability.

Key words: Callosobruchus maculatus, small heat shock protein(sHSP), temperature stress, mRNA expression level, real-time quantitative PCR(RT-qPCR)

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