棉铃虫表皮蛋白基因的克隆、序列及表达分析

Cloning,Sequence and Expression Analysis of Cuticle Protein Gene of Helicoverpa armigera

昆虫表皮蛋白能帮助昆虫抵御外界不良环境,在昆虫的生长发育中起重要作用。基于棉铃虫Helicoverpa armigera(Hübner)转录组数据信息克隆获得表皮蛋白基因的c DNA序列,通过Real-time PCR探讨棉铃虫各龄期和组织表皮蛋白的相对表达量。在棉铃虫体内得到HACPFL67 c DNA开放阅读框(KP072002)612 bp,命名为HACPFL67,编码204个氨基酸,分子量约为20.892 k Da,等电点7.275,序列包含有表皮蛋白的保守序列,与鳞翅目夜蛾科昆虫烟芽夜蛾Heliothis virescens和斜纹夜蛾Spodoptera litura等亲缘关系较近,与烟芽夜蛾氨基酸序列相似性高达83%。棉铃虫HACPFL67在不同发育阶段和组织中表达存在很大差异,其中4龄幼虫在各个发育阶段表达最高。综上所述,棉铃虫HACPFL67相对表达量在不同组织和发育时期存在一定差异。

Insect cuticle protein can help insects resist adverse environmental conditions and play an important role in the growth and development of insects. The c DNA sequence of the cuticle protein gene was cloned based on the information of the transcriptome of Helicoverpa armigera（Hübner） and the relative expression levels of the cuticle protein at different ages and tissues were investigated by Real-time PCR. A 612 bp HAPFL67 c DNA open reading frame（KP072002） was obtained in the bollworm, named HACPFL67, encoding 204 amino acids with a molecular weight of approximately 20.892 k Da and an isoelectric point of 7.275. The sequence contains a conserved sequence of cuticle protein, and is linked to Lepidoptera. Noctuidae Heliothis virescens and Spodoptera litura are closely related, among which the amino acid sequence similarity of the tobacco budworm is above 83%. The expression of HACPFL67 in cotton bollworms at different developmental stages and tissues was very different, and the fourth-instar larvae had the highest expression at various developmental stages. In summary, the relative expression levels of HACPFL67 in cotton bollworms indicate differences in different tissues and developmental stages.