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-  2019 

黄野螟精氨酸激酶基因的鉴定与表达分析
Identification and expression analysis of arginine kinase gene in Heortia vitessoides

Keywords: 黄野螟 精氨酸激酶 RT-qPCR 基因表达谱 温度胁迫
Heortia vitessoides arginine kinase RT-qPCR gene expression pattern temperature stress

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Abstract:

为了探索精氨酸激酶(arginine kinase,AK)在昆虫中的功能,经筛选黄野螟(Heortia vitessoides)成虫转录组数据库获得黄野螟精氨酸激酶基因,命名为HvAK(GenBank:MH794282),对其进行生物信息学、各虫态和幼虫组织的定量分析,并通过检测HvAK在高温及低温胁迫条件下的表达水平,揭示HvAK基因在黄野螟生长发育中的作用。结果显示:该序列开放阅读框长1 068 bp,共编码355个氨基酸;同源比对结果表明HvAK与家蚕(Bombyx mori)AK基因同源性最高,为94.10%;实时荧光定量PCR结果显示,黄野螟在所检测的各幼虫组织和发育阶段均有表达,其中在头部的表达量较高,在5龄幼虫阶段到达表达量最高峰;此外,高、低温均诱导HvAK表达量上调,推测HvAK基因在黄野螟生长发育过程中可能参与抵御外界不良环境。
The arginine kinase in Heortia vitessoides was obtained by screening the adult H.vitessoides transcriptome database and named HvAK (GenBank:MH794282).Bioinformatic analyses of HvAK and expression of HvAK at various developmental stages and larval tissues were performed.The role of HvAK gene in the growth and development of H. vitessoides was studied by detecting the expression level of HvAK under high temperature and low temperature stress.The results showed that the HvAK cDNA contains an open reading frame with 1 068 bp,encoding a protein with 355 amino acid.Result of homologous alignment showed that the highest homology between HvAK and AK gene of Bombyx mori was 94.10%.The results of real-time quantitative PCR (RT-qPCR) showed that HvAK was expressed in all larval tissues and developmental stages tested,with the highest expression of HvAK in the head and in the 5th instar larva.Both high and low temperature induced upregulation of HvAK expression,indicating that HvAK gene may be involved in resisting adverse external environment during H. vitessoides growth and development

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