棉铃虫BR‑C Z1基因的克隆及表达谱分析

Cloning and Expression Profiling of BR‑C Z1 Gene in Helicoverpa armigera

广泛锌指复合物(BR‐C)在棉铃虫Helicoverpa armigera的生长发育过程中有重要作用。基于转录组数据通过PCR克隆获得棉铃虫BR‑C Z1(HaBR‑C Z1)基因序列,利用生物信息学分析其氨基酸序列,利用qRT-PCR分析基因的表达规律,以及2-十三烷酮(2‐TD)处理后基因的表达情况。结果显示,HaBR‑C Z1基因的开放阅读框长1284 bp,编码427个氨基酸,预测编码蛋白的分子量和等电点分别为47.41 kD和7.72。系统进化树显示,HaBR‐C Z1与帝王斑蝶Danaus plexippus plexippus的亲缘关系最近。HaBR‑C Z1基因在预蛹期和6龄幼虫肠组织中的相对表达量最高。20 mg·g^(−1)浓度2-TD处理后的相对表达量在6 h达到峰值,为对照组的24.7倍。本研究明确了HaBR‑C Z1基因的表达规律及其对2-TD的响应,为深入了解棉铃虫生长发育机制奠定了基础,并为害虫防治提供依据。

Broad complex(BR‐C)has an important role in the growth and development of Helicoverpa armigera.In this study,the cDNA sequence of BR‑C Z1 gene was cloned from the midgut of H.armigera larvae based on previous transcrip‐tome data,and its amino acid sequence and protein structure were predicted.The fusion protein was expressed by using a prokaryotic expression system and verified by Western blot.Real-time quantitative PCR was used to detect the temporal and spatial expression profiles of BR‑C Z1 gene in H.armigera(HaBR‑C Z1),and the expression of BR‑C Z1 gene in the 6th instar larvae was detected after 2-tridecanone treatment.The results showed that the open reading frame of HaBR‑C Z1 gene was 1284 bp in length,encoding 427 amino acids.The predicted molecular weight and isoelectric point of HaBR‐C Z1 were 47.41 kD and 7.72,respectively.Phylogenetic analysis showed that HaBR‐C Z1 was closely related to the one of Danaus plexippus plexippus.The relative expression level of HaBR‑C Z1 gene was the highest at the pupal stage and in the midgut of the pre-pupa instar larvae.By contrast,the expression of HaBR‑C Z1 gene treatment with 20 mg·g^(−1) of 2-TD peaked(24.7 times as high as that of the control)at 6 h,and then gradually decreased.These results suggested that HaBR‐C Z1 might be involved in the growth and development of H.armigera,and respond to 2-TD stimulation.This study contributes to clarifying the expression profiling of HaBR‑C Z1 gene and provides a basis for understanding the growth and development mechanism of H.armigera in‐depth and pest control.