四纹豆象小分子量热激蛋白基因的鉴定与表达分析

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

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(Gen Bank编号FK669241.1)。采用实时荧光定量PCR(RT-q PCR)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 m RNA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。

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 to30 ℃ 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 upregulatedit stronger,that of Rec was slightly higher than that of CK,but not significantly. It isconsidered that there is no correlationship between shsp27 expression and high/low temperature adaptability.