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文冠果种子高表达XsKCS7基因的克隆和酵母表达功能鉴定

Cloning and Yeast Expression Function Identification of High Expression XsKCS7 Gene in Xanthoceras sorbifolium Seed

    摘要
  • 摘要:
    目的 探究调控文冠果种子神经酸合成关键基因。
    方法 本研究根据参考基因组联合转录组分析文冠果3-酮酯酰-CoA合酶（3-ketoacyl-CoA synthase，KCS）基因家族在不同发育时期种子中的表达模式； 通过RT-PCR扩增文冠果XsKCS7基因并进行生物信息学分析；XsKCS7基因异源转化酿酒酵母鉴定基因功能。
    结果 文冠果XsKCS7基因在不同发育时期种子中的表达量远高于其他KCS基因；克隆XsKCS7基因，生物信息学分析显示，XsKCS7基因的开放阅读框为1 512 bp，编码503个氨基酸，含有典型的KCS家族保守基序“GMGCSA”、“FGNTSSSS”以及“GSGFKCNSAVW”，与橡胶树KCS的亲缘性关系最近，为67.62%； XsKCS7异源转化酿酒酵母鉴定其具有调控芥酸和神经酸合成的功能。
    结论 XsKCS7基因确为调控文冠果种子芥酸和神经酸合成的关键基因。

     

    Abstract:
    Objective To explore the key genes of regulating nervonic acid biosynthesis in Xanthoceras sorbifolium seeds.
    Method This study analyzed the expression pattern of the 3-ketoesteryl-CoA synthase (3-ketoacyl-CoA synthase, KCS) gene family in seeds at different developmental periods according to the combined transcriptome of the reference genome. Gene XsKCS7 was cloned by RT-PCR, and was analyzed by Bioinformatics analysis of XsKCS7 protein. And the XsKCS7 gene was transformed into Saccharomyces cerevisiae to identify the gene function.
    Result The expression level of XsKCS7 gene in seeds of X. sorbifolium was much higher than other KCS genes. The open reading frame of XsKCS7 was 1,512 bp, encoding 503 amino acids, havd three typical conserved domains of KCS family "GMGCSA", "FGNTSSSS" and "GSGFKCNSAVW", and a close relationship with the homologous gene of Hevea brasiliensis,at 67.62%. XsKCS7 heterologous transformation of Saccharomyces cerevisiae regulated the synthesis of erucic acid and nervonic acid.
    Conclusion : XsKCS7 gene is a key gene that regulates the synthesis of erucic acid and nervonic acid in seeds of X. sorbifolium.

     

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