克氏原螯虾PcDsx基因的克隆及表达分析
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国家自然科学基金青年基金项目(31501858) ;中央高校基本科研业务费专项(2662020SCPY004);湖北省第二批现代农业产业技术体系“稻田综合种养创新团队”;中国水产科学研究院基本科研业务费专项(2019HY-JC)


Molecular cloning and expression analysis of PcDsx in Procambarus clarkii
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    摘要:

    为阐明克氏原螯虾(Procambarus clarki) doublesex(PcDsx)的功能,采用RACE技术克隆获得PcDs cDNA序列,利用qRTPCR检测该基因的表达情况。生物信息学分析结果显示,该基因cDNA全长1 584 bp,包括243 bp 5′ UTR、765 bp ORF(编码254 aa)和576 bp 3′ UTR;PcDsx蛋白包含1个保守的DM结构域,与东方刺龙虾(Sagmariasus verreauxi)SvDsx的DM结构域相似性较高。基因表达分析结果显示,PcDsx广泛表达于成年克氏原螯虾的各组织中,其中克氏原螯虾触角腺中该基因的相对表达量最高,性腺和肌肉中的相对表达量也较高,并且发现该基因在成年雌虾多种组织中的表达与相应雄虾组织中的表达存在显著性差异;克氏原螯虾早期发育不同时期表达分析结果表明,PcDsx的表达水平在出膜后3 d达到一个高峰,而幼虾中PcDsx的最高表达分别出现在出膜后41 d和115 d,此外,该基因在出膜后期雌雄幼虾中的表达亦表现出显著性差异。

    Abstract:

    To elucidate the function of the doublesex(PcDsx)gene in Procambarus clarkii ,the PcDsx cDNA sequence was obtained by the rapid amplification of cDNA ends (RACE) and the expression of PcDsx was determined by quantitative realtime PCR (qRT-PCR). The results of bioinformatics analysis showed that the fulllength PcDsx cDNA was 1 584 bp,with a 243 bp 5′ untranslated region,a 765 bp open reading frame (ORF) (coded 254 aa) and a 576 bp 3′ untranslated region. The deduced protein of PcDsx was 254 aa and the predicted PcDsx protein was found to contain a conserved DM domain. Phylogenetic analysis revealed that the PcDsx DM domain share high similarity to the DM domain of Sagmariasus verreauxi doublesex. The gene expression analysis showed that PcDsx was widely expressed in various tissues of the adult crayfish,highest in the antennary glands,followed by the muscle and gonads. The expression of PcDsx in various adult female tissues was significantly different from that in adult male tissues. During the early development of P. clarkii ,the expression level of PcDsx reached a peak at 3 day after hatching,while the highest expression of PcDsx in male and female juveniles appeared at 41 and 115 day after hatching,respectively. Besides,the expression of PcDsx also showed significant differences between male and female juveniles.

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石林林,陈家豪,石瑞雪,费佳敏,张龙,李艳和.克氏原螯虾PcDsx基因的克隆及表达分析[J].华中农业大学学报,2021,(1):129-136

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  • 收稿日期:2020-05-18
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  • 在线发布日期: 2021-02-10
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