丽江猪HMOX2基因扩增、序列分析及组织表达研究

doi:10.16431/j.cnki.1671-7236.2025.05.026

摘要/Abstract

摘要： 【目的】旨在扩增并探究丽江猪血红素加氧酶2(HMOX2)基因编码区(CDS)序列特征，分析在丽江猪不同生长阶段、不同组织中的表达情况，为利用HMOX2基因进行分子标记辅助育种奠定基础。【方法】利用PCR扩增丽江猪HMOX2基因序列，采用生物信息学软件分析其理化性质和密码子偏好性，预测其蛋白结构；采用实时荧光定量PCR检测HMOX2基因在2月龄丽江猪心脏、肝脏、脾脏、肺脏、肾脏和背最长肌，以及2、4、6月龄丽江猪皮下脂肪中的表达水平，并对HMOX2基因在2月龄丽江猪和杜洛克猪皮下脂肪中的表达水平进行比较分析。【结果】丽江猪HMOX2基因CDS序列长为951 bp，编码316个氨基酸，密码子偏好以C或G结尾。丽江猪HMOX2基因与黄牛、水牛、山羊和绵羊的相似性较高(90.1%~91.2%)，与原鸡的相似性最低(71.3%)。在6个地方猪种中，除八眉猪和荣昌猪外，丽江猪HMOX2基因与中国地方猪相似性为100%，与5个外种猪的相似性均为99.8%。与杜洛克猪相比，丽江猪HMOX2基因CDS区存在两处碱基同义突变。HMOX2蛋白属于稳定的跨膜亲水性蛋白，无信号肽，具有1个跨膜结构，主要在细胞质中发挥作用，含21个磷酸化位点和33个糖基化位点。二级结构主要由α-螺旋(54.43%)和无规则卷曲(42.00%)组成。蛋白互作网络分析表明，HMOX2蛋白可能与FECH、HMOX1等10个蛋白互作，主要在脂质代谢中发挥作用。实时荧光定量PCR结果表明，HMOX2基因在丽江猪心脏、肝脏、脾脏、肺脏、肾脏、背最长肌和皮下脂肪中均有表达,且在皮下脂肪中随月龄增长表达量增加。与杜洛克猪相比，丽江猪HMOX2基因在皮下脂肪中的表达量显著高于杜洛克猪(P＜0.05)。【结论】本研究成功扩增了丽江猪HMOX2基因并分析了其分子特征，其在肺脏、肝脏和皮下脂肪等7个组织中均有表达，在皮下脂肪组织中随月龄增长表达量显著增加。研究结果可为进一步探究HMOX2基因在丽江猪脂肪沉积中的作用奠定基础。

关键词: 丽江猪; 血红素加氧酶2基因; 序列分析; 组织表达

Abstract: 【Objective】 This study was aimed to amplify and explore the coding sequence (CDS) sequence characteristics of heme oxygenase 2 (HMOX2) gene in Lijiang pigs, and analyze its expression in different growth stages and tissues of Lijiang pigs,establishing a foundation for molecular marker-assisted breeding utilizing HMOX2 gene.【Method】 The HMOX2 gene sequence of Lijiang pigs was amplified by PCR.The physicochemical properties,codon preferences,and predicted protein structure were analyzed using bioinformatics software.The expression of HMOX2 gene were detected using Real-time quantitative PCR in heart,liver,spleen,lung,kidneys,and longissimus dorsi muscle of 2-month-old Lijiang pigs,as well as in the subcutaneous fat of 2-,4-,and 6-month old Lijiang pigs,and compare the expression of HMOX2 gene in subcutaneous fat of 2-month-old Lijiang pigs and Duroc pigs.【Result】 The CDS of HMOX2 gene in Lijiang pigs was found to be 951 bp long,encoding 316 amino acids,and the codon prefers to end with C or G.The HMOX2 gene in Lijiang pigs showed high similarity to Bos taurus, Bos indcus, Capra hircus and Ovis aries (90.1%-91.2%),and the lowest similarity to Gallus gallus (71.3%).Among six local pig breeds,except Bamei pigs and Rongchang pigs, HMOX2 gene in Lijiang pigs showed 100% similarity to local pigs,and the similarity with five exotic pigs was 99.8%.Two base synonymous mutations were present in the CDS region of HMOX2 gene in Lijiang pigs compared with Duroc pigs. HMOX2 protein were determined to be stable transmembrane hydrophilic proteins without a signal peptide,featuring one transmembrane structure,primarily functioning in cytoplasm,and containing 21 phosphorylation and 33 glycosylation sites.The secondary structure of HMOX2 protein was predominantly composed of alpha helix (54.43%) and random coil (42.00%).Protein interaction network analysis demonstrated that HMOX2 protein might interact with 10 proteins,including FECH and HMOX1,primarily playing a role in lipid metabolism.Real-time quantification PCR results indicated that HMOX2 gene was expressed in heart,liver,spleen,lung,kidney,longissimus dorsi muscle,and subcutaneous fat of Lijiang pigs,and the expression increased with the increase of age in subcutaneous fat.Compared with Duroc pigs,the expression of HMOX2 gene in subcutaneous fat of Lijiang pigs was significantly higher than that of Duroc pigs (P＜0.05).【Conclusion】 HMOX2 gene in Lijiang pigs was successfully amplified,and its molecular characteristics were elucidated.It was expressed in seven tissues such as lung,liver,and subcutaneous fat,and its expression in subcutaneous fat increased significantly with the increase in monthly age.These results provided a foundation for further investigation into the role of HMOX2 gene in fat deposition in Lijiang pigs.

Key words: Lijiang pigs; HMOX2 gene; sequence analysis; tissue expression

中图分类号:

S828

李天秀, 李昕鹏, 董新星, 兰国湘, 严达伟, 朱家位. 丽江猪HMOX2基因扩增、序列分析及组织表达研究[J]. 中国畜牧兽医, 2025, 52(5): 2219-2231.

LI Tianxiu, LI Xinpeng, DONG Xinxing, LAN Guoxiang, YAN Dawei, ZHU Jiawei. Amplification,Sequence Analysis and Tissue Expression Study of HMOX2 Gene in Lijiang Pigs[J]. China Animal Husbandry and Veterinary Medicine, 2025, 52(5): 2219-2231.

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