• 中文核心期刊
  • CSCD来源期刊
  • 中国科技核心期刊
  • CA、CABI、ZR收录期刊

克氏原螯虾受鲶爱德华氏菌刺激后Pc-crustin 6基因的表达分析

Gene Cloning and Expression of Crustin 6 in Procambarus clarkii

  • 摘要:
      目的  为了后续深入研究克氏原螯虾甲壳肽基因的功能。
      方法  以Pc-crustin 6甲壳肽基因作为研究对象,通过设计特异性引物进行PCR扩增,利用生物信息学软件进行氨基酸序列比对与系统进化树分析,探讨Pc-Crustin 6的分子特性;并利用qRT-PCR技术分析目的基因在不同组织中的表达水平,检测鲶爱德华氏菌刺激后基因的表达模式。
      结果  Pc-crustin 6的cDNA序列全长为465 bp,开放阅读框包括384 bp,编码127个氨基酸残基;在分子的N端包含一个由25个氨基酸残基组成的信号肽序列,C端包含一个由8个保守的半胱氨酸残基构成的WAP结构域。Pc-crustin 6在被检测的5个组织中均存在一定的表达,其中,鳃组织的相对表达量最高。鲶爱德华氏菌刺激后,Pc-crustin 6在血细胞、肝胰腺、鳃、肠组织中均表现出了明显的上调趋势。
      结论  机体在受到鲶爱德华氏菌刺激后,Pc-crustin 6相对表达量发生改变,推测其在小龙虾抗鲶爱德华氏菌过程中发挥了作用。

     

    Abstract:
      Objective  Functions of crustacean gene of Procambarus clarkia were studied.
      Method  Pc-crustin6 was targeted for this study with specifically designed primers for PCR. Amino acids alignment and phylogenetic tree analysis were performed with bioinformatics software to decipher the molecular properties of Pc-crustin 6. Expressions and distributions of Pc-crustin 6 in different tissues were analyzed by qRT-PCR. The expression after an Edwardsiella ictaluri challenge was used to determine the disease resistance associated with the gene.
      Result  The full length Pc-crustin 6 cDNA was 465 bp with an open reading frame of 384 bp and encoding 127 amino acids. The N terminal of the molecule contained a signal peptide of 25 amino acids, and the C terminal a WAP domain of 8 conserved cysteine residues. Pc-crustin 6 was expressed to varied extents in 5 organs including the gills, which was the highest among them. After an artificial exposure to E. ictaluri, the expressions of Pc-crustin 6 in the hemocytes, hepatopancreas, gills, and intestines of P. clarkia were significantly up-regulated.
      Conclusion  The Pc-crustin 6 expression in organs of P. clarkia was significantly altered after being stimulated by the induction of E. ictaluri. It indicated that the gene was involved in the pathogenic resistance of P. clarkia.

     

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