陆川猪CSRP3基因克隆及其组织表达差异分析

Cloning and differential expression analysis of CSRP3 gene in Luchuan pig

  • 摘要: 【目的】克隆陆川猪的富含半胱氨酸和甘氨酸蛋白3(CSRP3)基因,并对其进行生物信息学和组织表达谱分析,为研究CSRP3基因在陆川猪肌肉生长过程中的作用机制打下基础。【方法】根据NCBI已公布的野猪CSRP3基因序列设计特异性定量引物和克隆引物,运用RT-PCR克隆CSRP3基因并进行生物信息学在线分析,采用实时荧光定量PCR检测CSRP3基因在陆川猪各组织中的表达差异。【结果】陆川猪CSRP3基因编码区(CDS)全长854 bp,编码194个氨基酸残基,与NCBI已公布的野猪CSRP3基因CDS序列相比存在5处同义碱基突变,且陆川猪与野猪氨基酸序列相似性达100.0%;陆川猪CSRP3基因的编码蛋白分子量为20935.82 Da,分子式为C964H1404N262O274S19,理论等电点(pI)为8.89,不稳定系数为39.11,表明其是偏碱性的稳定蛋白;CSRP3蛋白二级结构主要由无规则卷曲构成,三级结构可能有一种模型;陆川猪CSRP3蛋白除有多个磷酸化位点之外,无跨膜结构和信号肽;陆川猪CSRP3基因在心脏中表达量最高,背最长肌次之,在肝脏中的表达量最低。【结论】CSRP3基因在陆川猪心脏中表达量最高,背最长肌次之且明显高于其他组织,说明该基因可能对肌肉生长有一定影响。

     

    Abstract: 【Objective】The cysteine and glycine rich protein 3(CSRP3)of Luchuan pig was cloned and the bioinformatics and tissue expression profile were analyzed,so as to lay foundation for studying the role of CSRP3 gene in muscle growth of Luchuan pigs.【Method】According to the sequence of wild boar CSRP3 gene published on NCBI,specific quantitative primers and cloning primers were designed,CSRP3 gene was cloned by RT-PCR,and bioformatics analysis was conducted online.Finally,the expression difference of CSRP3 gene in various tissues of Luchuan pig was detected by qPT-PCR.【Result】The CDS region of Luchuan pig CSRP3 gene was 854 bp in length,encoding 194 amino acids.Compared with the wild boar CSRP3 gene CDS region published on NCBI,there were 5 synonymous base mutations.The amino acid sequence was the same and its similarity was 100.0%between Luchuan pig and wild boar.Molecular weight of the protein encoded by the CSRP3 gene of Luchuan pig was 20935.82 Da,and its molecular formula was C964H1404N262O274S19,the theoretical isoelectric point was 8.89 and the instability coefficient was 39.11,which indicated that it was a stable protein with alkalinity.The secondary structure of CSRP3 gene was mainly composed of random coils and there might be a model of tertiary structure.Luchuan pig CSRP3 protein had no transmembrane structure and signal peptide except for multiple phosphorylation sites.Luchuan pig CSRP3 gene had the highest expression in the heart,followed by muscle tissue.The expression in the liver was the lowest.【Conclusion】The expression of CSRP3 gene is highest in the heart of Luchuan pigs,followed by muscle.And the expression in heart and muscle are significantly higher than that in other tissues,indicating that this gene may have a certain effect on muscle growth.

     

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