荔枝粗胫翠尺蛾卵黄原蛋白基因及其受体基因的克隆与生物信息学分析

    Cloning and Bioinformatics Analysis of Vitellogenin Genes and Its Receptor Genes in Thalassodes immissaria

    • 摘要:
      目的 卵黄原蛋白(Vitellogenin,Vg)通过其受体(Vitellogenin Receptor,VgR)介导的内吞作用,为卵母细胞的卵黄发生提供必需营养,二者在昆虫生殖调控中发挥关键作用。旨在探究荔枝重要食叶害虫粗胫翠尺蛾(Thalassodes immissariaVg及其受体VgR基因的生物信息学特征和在荔枝粗胫翠尺蛾生殖调控中的功能。
      方法 根据荔枝粗胫翠尺蛾转录组信息,利用PCR技术克隆TiVgTiVgR的全长cDNA序列,并通过ORF Finder、SignalP 5.0、ProtParam、SMART、TMHMM 2.0、MEGA v5.05等软件对其进行生物信息学分析;根据荔枝粗胫翠尺蛾卵、幼虫、蛹和成虫转录组数据和qRT-PCR技术,分析TiVgTiVgR在不同发育阶段和不同组织的mRNA表达模式。
      结果 TiVgTiVgR的开放阅读框(ORF)长度分别为5 343和5 250 bp,分别编码1 780和1 749个氨基酸残基的多肽链,预测分子量分别为202.403和196.248 kDa;二者均为亲水性蛋白,亲水性指数分别为-0.681和-0.329。信号肽预测结果显示,TiVg和TiVgR的N端分别存在1个由16和14个氨基酸残基组成的信号肽区域。系统发育分析表明,TiVg与冬尺蠖(Operophtera brumata)的Vg亲缘关系最近,TiVgR与棉铃虫(Helicoverpa armigera)的VgR亲缘关系最近。以TPM值为表达量的表达模式结果显示,TiVg在蛹期开始表达,雌成虫期显著高表达,TiVgR在雌成虫期显著高表达,卵期微弱表达。qRT-PCR结果表明,TiVg在雌成虫脂肪体相对高表达,TiVgR在雌成虫卵巢相对高表达。
      结论 成功克隆荔枝粗胫翠尺蛾的VgVgR基因,二者含有VgVgR家族保守的元件,且不同发育阶段及组织的表达模式结果表明,TiVgTiVgR可能参与荔枝粗胫翠尺蛾生殖调控的生理过程。研究结果可为荔枝粗胫翠尺蛾生殖调控机制的研究奠定理论基础。

       

      Abstract:
      Objective Vitellogenin (Vg) and its receptor (VgR) play key roles in insect reproduction by mediating nutrient transport during oocyte vitellogenesis.This study aimed to clone Vg and VgR from the Thalassodes immissaria, a notorious defoliator in lichee, and analyzed their bioinformatics characteristics and expression patterns across developmental stages to infer their roles in reproductive regulation.
      Method Using transcriptome data of T.immissaria, the full-length cDNA sequences of TiVg and TiVgR were cloned via PCR.Bioinformatics tools, such as ORF Finder, SignalP 5.0, ProtParam, SMART, TMHMM 2.0, and MEGA v5.05, were used for sequence analysis.Expression profiles of TiVg and TiVgR across different developmental stages and different tissues were quantified using transcriptomic TPM values and qRT-PCR.
      Result The open reading frames (ORFs) of TiVg and TiVgR span 5 343 and 5 250 bp respectively, encoding polypeptides of 1 780 and 1 749 amino acids, with predicted molecular weights of 202.403 and 196.248 kDa.Both TiVg and TiVgR are water-soluble, exhibiting hydropathicity index of-0.681 and-0.329, respectively.The N-termini of TiVg and TiVgR contain signal peptide regions composed of 16 and 14 amino acid residues, respectively.The phylogenetic analysis suggested that TiVg clusters within the Operophtera brumata ObVg clade, while TiVgR aligns closely with Helicoverpa armigera HaVgR orthologs.The expression pattern results with TPM value showed that TiVg expression initiated at the pupal stage and peaked in adult females, whereas TiVgR exhibited maximal expression in adult females with minimal levels detected in eggs.Tissue expression pattern suggested that TiVg was highly expressed in fat body of adult females and TiVgR was highly expressed in ovary of adult females, respectively.
      Conclusion The study successfully cloned Vg and VgR from T.immissaria.The conserved motifs and the expression patterns in both developmental stage and tissues suggest their potential roles in reproductive regulation.These findings provide a foundation for understanding reproductive mechanisms in T.immissaria.

       

    /

    返回文章
    返回