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      2. 克氏原螯蝦Sex-lethal基因的克隆與表達分析
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        華中農業大學

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        中央高?;究蒲袠I務費專項資金資助項目(2662020SCPY004);國家自然科學基金青年基金項目(31501858) ;湖北省第二批現代農業產業技術體系“稻田綜合種養創新團隊”;中國水產科學研究院基本科研業務費專項(2019HY-JC)


        Cloning and expression analyses of Sex-lethal gene in Procambarus clarkii
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          摘要:

          為探究克氏原螯蝦Sex-lethal基因的功能與分子機制,采用RACE(rapid amplification of cDNA ends)技術克隆獲得4個克氏原螯蝦Sxl cDNA序列,利用熒光定量PCR技術檢測其在不同組織以及早期發育時期的表達情況。生物信息學分析發現, PcSxlβ和PcSxlδ比PcSxlγ多了一段77 bp的堿基序列,而PcSxlβ和δ的預測所得的氨基酸序列較PcSxlγ短??耸显r的Sxl氨基酸序列與紅螯螯蝦的序列相似性較高(75.76%)。保守結構域分析顯示,這4種轉錄異構體序列預測的氨基酸序列都具有相同的2個高度保守的RRM結構域。表達分析結果顯示,PcSxl在成年雄性克氏原螯蝦的觸角腺和鰓中表達量較高,在成年雌性中則是中腸和前腸的表達量較高,雌性卵巢中的表達量要顯著高于雄性精巢。在檢測的早期發育時期中,無節幼體時期的表達量最高,在蚤狀幼體期表達量卻出現下降,但在出膜后第1天表達量又有所升高,隨后整體則呈現出逐漸下降的趨勢。以上結果表明克氏原螯蝦PcSxl與其性別分化有關。

          Abstract:

          In order to preliminarily explore the function and molecular mechanism of Sex-lethal gene in Procambarus clarkii, four Sxl cDNA sequences from P. clarkii were cloned using rapid amplification of cDNA ends, and their expression in different tissues and early developmental stages were detected by quantitative fluorescence PCR. Bioinformatics analyses revealed that PcSxlβ and PcSxlδ had a 77 bp more base sequence than PcSxlγ, and the predicted amino acid sequence of PcSxlβ and PcSxlδ were shorter than that of PcSxlγ. The PcSxl sequences was highly similar to that of Cherax quadricarinatus (75.76%). The analysis of conservative domain showed that the amino acid sequences predicted by the four transcriptional isomers all had the same two highly conservative RRM domains. The expression analyses showed that PcSxl was highly expressed in the antennal glands and gills of adult males, and in the midgut and foregut of adult females, and was significantly higher in the female ovaries than the male testis. In the early developmental period of the crayfish, the expression was the highest in the period of nauplius stage, but decreased in the zoea stage; however, the expression level increased at the first day after hatching, and then showed a gradually decreasing trend as a whole. The results of this study suggested that PcSxl was associated with sexual differentiation in P. clarkii.

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        • 收稿日期:2020-06-04
        • 最后修改日期:2020-10-16
        • 錄用日期:2020-12-03
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