高温胁迫下桃小食心虫热激蛋白Hsp90和Hsp70基因的克隆及序列分析

摘要

桃小食心虫(Carposina sasakii Matsumura)是果树上重要的食心虫类害虫,热激蛋白Hsp90和Hsp70在昆虫抵御温度胁迫反应中具有重要作用。采用RT-PCR方法克隆获得了高温胁迫下桃小食心虫热激蛋白Hsp90和Hsp70基因cDNA部分序列,并对其进行分析,为深入揭示桃小食心虫对环境适应的分子机理提供理论依据。已获得的桃小食心虫热激蛋白Hsp90基因(GenBank登录号：KJ139642)序列长420bp,编码139个氨基酸残基,推导的氨基酸序列中含有热激蛋白90家族的一段序列为YSNKEIFLRE的特征序列,并且cDNA序列在N端具有Hsp90基因保守的ATPase结构,该序列与天蚕(Antheraea yamamai)和柞蚕(Antheraea pernyi)等昆虫的氨基酸序列一致性高达99%。已获得的Hsp70-1基因(GenBank登录号：KJ139643)和Hsp70-2基因(GenBank登录号：KJ139644)序列长均为305bp,编码101个氨基酸残基。 Hsp70-1基因推导的氨基酸序列与家蚕(Bombyx mori)的氨基酸序列一致性为94%,Hsp70-2基因推导的氨基酸序列与小菜蛾(Plutella xylostella)和烟草夜蛾(Manduca sexta)等昆虫的氨基酸序列一致性为96%。%Peach fruit moth,Carposina sasakii Matsumura is one of the main pests of the fruit trees. The heat shock protein 90 (Hsp90)and the heat shock protein70(Hsp70)widely exist in insects and play important role under temperature stress. In order to provide theoretical basis for exploring molecular mechanism of insect adaption to environment, partial-length cDNA encoding Hsp90 and Hsp70 from Carposina Sasakii were cloned by RT-PCR and the sequence was analyzed. Sequence analysis indicated that the partial cDNA of Hsp90 (GenBank accession number: KJ139642) was 420bp, encoding 139 amino acid residues. The deduced amino acid sequence contains an important Hsp90 family characteristic sequence(YSNKEIFLRE) and conservative ATPase structure in N-terminal domain, which shares 99% amino acid sequence identies with the Hsp90 genes from Antheraea yamamai and Antheraea pernyi. The partial cDNAs of Hsp70-1 (GenBank accession number: KJ139643) and Hsp70-2 (GenBank accession number: KJ139644) were all 305bp, encoding101 amino acid residues. The deduced amino acid sequence of Hsp70-1 shares 94% amino acid sequence identies with the Hsp90 genes from Bombyx mori, and the deduced amino acid sequence of Hsp70-2 shares 96% amino acid sequence identies with the Hsp90 genes from Plutella xylostella and Manduca sexta.