棉铃虫c-Jun氨基末端激酶基因的克隆、表达谱及对UV-A胁迫的响应

doi:10.16380/j.kcxb.2019.04.003

摘要/Abstract

摘要： 【目的】本研究旨在克隆棉铃虫Helicoverpa armigera c-Jun氨基末端激酶(c-Jun N-terminal kinase, JNK)基因，并对其进行序列和表达模式分析，探讨该基因在棉铃虫生长发育及响应UV胁迫方面的作用。【方法】利用RT-PCR与RACE技术克隆棉铃虫JNK基因，并利用生物信息学方法对其编码的氨基酸序列进行分析；采用实时荧光定量PCR技术检测其在棉铃虫不同发育阶段 (卵、1-6龄幼虫、蛹、雌雄成虫)、成虫不同组织(去除触角和复眼的头、胸、腹、触角、复眼、足、翅、中肠、卵巢)中及雌成虫在UV-A照射不同时间(0, 30, 60, 90, 120和150 min)下的相对表达量变化。【结果】克隆获得一个棉铃虫JNK基因并命名为HaJNK (GenBank登录号: MH719009)，其cDNA序列全长为2 431 bp，开放阅读框(ORF)长1 191 bp，编码396个氨基酸，编码蛋白质的相对分子量为45.01 kD，等电点为6.35，无跨膜结构，无信号肽。系统进化分析显示，棉铃虫HaJNK与其他昆虫JNK具有很高的同源性。发育阶段表达分析表明，HaJNK在棉铃虫卵期表达量最高；组织特异性分析显示该基因在成虫复眼、胸部及卵巢部位特异性表达。UV-A照射能诱导棉铃虫雌成虫体内HaJNK的表达，随着照射时间的延长，其表达量呈现先升高后降低的趋势，在照射60 min时表达量达到峰值。【结论】HaJNK在棉铃虫不同龄期、成虫不同组织和UV-A照射不同时间的雌成虫中差异表达，提示其在响应UVA胁迫的分子机制中具有重要意义。

关键词: 棉铃虫, c-Jun氨基末端激酶, 基因克隆, 表达分析, UV-A胁迫

Abstract: 【Aim】 This study aims to clone the c-Jun N-terminal kinase (JNK) gene and to analyze the gene sequence and mRNA expression profiles in Helicoverpa armigera, so as to explore the function of JNK gene in the growth and development of the moth and its responses to UV-A stress. 【Methods】 The full-length cDNA of the JNK gene of H. armigera was cloned and identified using RT-PCR and RACE technique. The putative amino acid sequence of the JNK gene was analyzed by bioinformatics methods. The expression levels of the JNK gene in different developmental stages (egg, 1st-6th instar larva, pupa, female and male adult), adult tissues (head with antennae and compound eyes removed, thorax, abdomen, antenna, compound eye, legs, wing, midgut and ovary) and female adults exposed to UV-A for various time periods (0, 30, 60, 90, 120 and 150 min) were detected by real-time quantitative PCR (RT-qPCR). 【Results】 A JNK gene was cloned from H. armigera, and named HaJNK (GenBank accession no.: MH719009), which is 2 431 bp in length and contains an open reading frame (ORF) of 1 191 bp, encoding 396 amino acid residues. The putative protein is 45.01 kD with an isoelectric point (pI) of 6.35, and has no transmembrane region and signal peptide. The phylogenic tree indicated that HaJNK shares a high homology with JNKs from other insect species. Developmental stage-specific mRNA expression profiling showed that HaJNK had the highest expression levels in the egg stage. Tissue-specific mRNA expression profiling showed that HaJNK was specifically expressed in the compound eye, thorax and ovary of adult. The mRNA expression of HaJNK in female adults was induced by UV-A stress, increasing firstly and then decreasing with the increase of exposure time, and reached the highest expression level at 60 min post exposure to UV-A. 【Conclusion】 This study indicates that the expression level of HaJNK varies at different developmental stages, adult tissues and female adults of H. armigera exposed to UV-A for different time, suggesting that HaJNK may play an important role in the response to UV-A stress.

Key words: Helicoverpa armigera, c-Jun N-terminal kinase, gene cloning, expression analysis, UV-A stress

刘小飞, 孟建玉, 赵晓超, 张长禹. 棉铃虫c-Jun氨基末端激酶基因的克隆、表达谱及对UV-A胁迫的响应[J]. 昆虫学报, 2019, 62(4): 407-417.

LIU Xiao-Fei, MENG Jian-Yu, ZHAO Xiao-Chao, ZHANG Chang-Yu. cDNA cloning and expression profiling of the c-Jun N-terminal kinase gene and its response to UV-A stress in Helicoverpa armigera (Lepidoptera: Noctuidae)[J]. Acta Entomologica Sinica, 2019, 62(4): 407-417.

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棉铃虫c-Jun氨基末端激酶基因的克隆、表达谱及对UV-A胁迫的响应

cDNA cloning and expression profiling of the c-Jun N-terminal kinase gene and its response to UV-A stress in Helicoverpa armigera (Lepidoptera: Noctuidae)

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