合作猪SIRT3基因克隆、生物信息学分析及组织表达研究

doi:10.16431/j.cnki.1671-7236.2024.04.007

摘要/Abstract

摘要： 【目的】克隆合作猪沉默信息调节因子3(silence information regulator 3,SIRT3)基因CDS区序列,并对其进行生物信息学分析,探究SIRT3基因在合作猪不同组织中的表达水平,为研究SIRT3基因功能奠定基础。【方法】试验选择3月龄合作猪公猪3头,采集心脏、睾丸、肺脏等组织,采用PCR扩增、Sanger测序等方法获取SIRT3基因CDS区序列,并通过Mega 7.0软件构建系统进化树;通过生物信息学在线软件分析SIRT3蛋白结构和功能;采用实时荧光定量PCR检测SIRT3基因在合作猪不同组织中的表达量。【结果】合作猪SIRT3基因CDS区长774 bp,共编码257个氨基酸。系统进化树结果显示,合作猪与家猪亲缘关系最近,与斑马鱼亲缘关系最远。SIRT3蛋白的分子质量为28.68 ku,理论等电点为5.81,不稳定系数为37.92,为稳定性亲水蛋白;该蛋白不存在跨膜区域,无信号肽,但存在2个低复杂度结构域,主要分布于内质网中;SIRT3蛋白的二级结构主要由α-螺旋和无规则卷曲构成,三级结构模型预测结果与二级结构基本一致。实时荧光定量PCR结果显示,SIRT3基因在合作猪不同组织中均有表达,在心脏中表达量最高,极显著高于其他组织(P＜0.01),其次是睾丸、肺脏、肝脏组织。【结论】本研究成功克隆合作猪SIRT3基因CDS区序列,探究了SIRT3基因在合作猪不同组织中的表达水平并初步分析了其编码蛋白的生物学功能,为进一步研究合作猪SIRT3基因功能提供了参考。

关键词: 合作猪; SIRT3基因; 克隆; 生物信息学分析; 组织表达

Abstract: 【Objective】 The CDS region of silence information regulator 3 (SIRT3) gene in Hezuo pigs was cloned and bioinformatic analysis was performed,and the expression level in different tissues of Hezuo pigs was explored,laying a foundation for the study of the function of SIRT3 gene.【Method】 Three 3-month-old Hezuo boars were selected to collect heart,testis,lung and other tissues.The CDS region sequence of SIRT3 gene was obtained by PCR amplification and Sanger sequencing,and the phylogenetic tree was constructed by Mega 7.0.Bioinformatic analysis of SIRT3 protein was performed by online software.The tissue expression of SIRT3 gene was detected by Real-time quantitative PCR.【Result】 The CDS region of SIRT3 gene was 774 bp,encoding 257 amino acids.Hezuo pig had the closest relationship with domestic pig,and had a distant relationship with zebrafish.The molecular weight of SIRT3 protein was 28.68 ku,the theoretical isoelectric point was 5.81,and the instability coefficient was 37.92,indicating that SIRT3 protein was a stable hydrophilic protein.SIRT3 protein had no transmembrane region and no signal peptide,but there were 2 low complexity region,which were mainly distributed in the endoplasmic reticulum.The secondary structure of SIRT3 protein was mainly alpha-helix and random coil,and the results predicted by the tertiary structure model were basically consistent with the secondary structure.Real-time quantitative PCR results showed that SIRT3 gene was expressed in different tissues of Hezuo pigs,and the expression level in heart was the highest,which was extremely significantly higher than that in other tissues (P＜0.01),followed by testis,lung and liver tissues.【Conclusion】 The CDS region sequence of SIRT3 gene in Hezuo pigs was successfully cloned,the expression level of SIRT3 gene in different tissues of Hezuo pigs was explored,and the biological function of its encoded protein was preliminatively analyzed,which provided a reference for further study of SIRT3 gene function in Hezuo pigs.

Key words: Hezuo pigs; SIRT3 gene; cloning; bioinformatics analysis; tissue expression

中图分类号:

S828.8⁺9

闫尊强, 梁毓豪, 宋科林, 滚双宝, 王鹏飞. 合作猪SIRT3基因克隆、生物信息学分析及组织表达研究[J]. 中国畜牧兽医, 2024, 51(4): 1390-1399.

YAN Zunqiang, LIANG Yuhao, SONG Kelin, GUN Shuangbao, WANG Pengfei. Cloning,Bioinformatics Analysis and Tissue Expression of SIRT3 Gene in Hezuo Pigs[J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(4): 1390-1399.

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