龟足碳酸酐酶(CA)基因家族的鉴定及表达分析

    Identification and expression analysis of carbonic anhydrase (CA) gene family in Capitulum mitella

    • 摘要: 碳酸酐酶(carbonic anhydrase, CA)是一类催化二氧化碳和水生成碳酸的酶。为探究CA在龟足壳板钙化过程中的作用,利用生物信息学的方法对龟足CA基因家族成员进行鉴定,分析其编码蛋白的理化性质、亚细胞定位、二级结构、三级结构、保守基序、染色体定位、跨膜结构预测及进化关系,并结合转录组分析龟足CA在不同发育时期的表达模式。结果表明:在龟足基因组中鉴定出10条CA基因家族成员,其分布在6条不同的染色体上。龟足CA基因家族成员编码蛋白的基本理化性质分析表明,蛋白氨基酸数目为108~318个,蛋白相对分子质量为12.10~36.22 kDa,等电点系数范围为5.09~10.68,且均为亲水蛋白;系统发育分析显示在龟足CA蛋白中,CmCA3、CmCA4和CmCA5聚为一支,CmCA6、CmCA7、CmCA8和CmCA9聚为另一支,而CmCA1、CmCA2和CmCA10则分别单独聚为一支;转录组数据分析显示CmCA7CmCA8CmCA2CmCA6CmCA10在无节幼虫时期表达量相对较低,CmCA5CmCA3分别在变态中期和稚体时期具有较高的表达量,CmCA4CmCA1在无节幼虫时期具有较高的表达量,CmCA9在金星幼虫5期表达量增加,随后显著下降。综上所述,研究结果为深入解析CA基因在龟足壳板钙化过程中的调控机制提供理论基础。

       

      Abstract: Carbonic anhydrase(CA)is an enzyme that reaction between carbon dioxide and water to produce carbonic acid. To investigate the function of CA gene family in the calcification process of Capitulum mitella, this study identified members of the CmCA gene family using bioinformatics methods, and analyzed their physicochemical properties, subcellular localization, secondary and tertiary structure, conserved motifs, chromosomal localization, transmembrane structure prediction and evolutionary relationships, and combined with transcriptome analysis of the expression pattern of CmCA in different developmental stages. The findings indicated that 10 CA gene family members were identified in the C.mitella genome, which were distributed on six different chromosomes. Physicochemical property analyses showed that the number of amino acids ranged from 108 to 318, with molecular weights between 12.10 kDa and 36.22 kDa and isoelectric points ranging from 5.09 to 10.68, and all of them were hydrophilic proteins. Phylogenetic analysis showed that CmCA3, CmCA4, and CmCA5 clustered into one group, while CmCA6, CmCA7, CmCA8, and CmCA9 formed another group. Meanwhile, CmCA1, CmCA2, and CmCA10 each formed independent group. Transcriptome analysis revealed that CmCA7, CmCA8, CmCA2, CmCA6, and CmCA10 were expressed at relatively low levels during the nauplius stage, while CmCA5 and CmCA3 showed high expression expressed during the metamorphosis mid-stage and juvenile stage, CmCA4 and CmCA1 exhibited high expression during the nauplius stage, while CmCA9 showed increased expression at the cyprid stage, followed by a significant decrease. In summary, the results of this study provide a theoretical basis for further understanding the regulatory mechanism of CA genes in the calcification process of C.mitella.

       

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