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2025年5月6日 星期二
农业生物技术学报  2019, Vol. 27 Issue (3): 441-448    DOI: 10.3969/j.issn.1674-7968.2019.03.007
  研究论文与报告 本期目录 | 过刊浏览 | 高级检索 |
藏绵羊HMOX1基因表达及其多态性与低氧适应性的关联分析
何建文1,2, 张伟1, 刘秀1,*, 李少斌1, 王继卿1, 胡江1, 罗玉柱1,*
1 甘肃农业大学 动物科学技术学院/甘肃省草食动物生物技术重点实验室/甘肃省牛羊基因改良工程实验室,兰州 730070;
2 甘肃农业职业技术学院,兰州 730020
HMOX1 Gene Expression and Assiciation Between Its Polymorphism and Hypoxia Adaptation in Tibetan Sheep (Ovis aries)
HE Jian-Wen1,2, ZHANG Wei1, LIU Xiu1,*, LI Shao-Bin1, WANG Ji-Qing1, HU Jiang1, LUO Yu-Zhu1,*
1 College of Animal Science and Technology/Gansu Key Laboratory of Herbivorous Animal Biotechnology/Gansu Engineering Laboratory of Genetic Improvement in Ruminants, Gansu Agricultural University, Lanzhou 730070, China;
2 GanSu Agriculture Technology College, Lanzhou 730020, China
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摘要 血红素加氧酶1(heme oxygenase 1, HMOX1)是血红素加氧酶家族的应激诱导型酶,在机体低氧胁迫代谢调控过程中扮演着重要角色。藏绵羊(Ovis aries)在长期进化过程中形成了独特的低氧适应策略来维持自身的氧平衡。为了探讨HMOX1基因的组织表达及其第三外显子(Exon3)区域多态性与藏绵羊高原低氧适应的关系,以寻找与藏绵羊低氧适应相关的分子标记,本研究以不同海拔地区的藏绵羊和湖羊(对照组)为研究对象,利用qRT-PCR及PCR-单链构象多态性(PCR-single-strand conformation polymorphism, PCR-SSCP)方法检测HMOX1基因的组织表达,以及Exon3区域多态位点,分析不同基因型及等位基因与绵羊血液指标的相关性,以及SNPs与绵羊血液指标的关联性。结果表明,HMOX1基因在藏绵羊和湖羊各组织中均有表达。其中同一品种内,HMOX1在藏绵羊脾脏中高表达(P<0.01),其次在肝、肾脏和背最长肌中表达量也较高(P<0.05);而在湖羊中,HMOX1在肺脏和脂肪中高表达(P<0.01),其次在脾脏、肾脏及肝脏中的表达量也较高(P<0.05)。2个品种间同一组织表达比较结果显示,HMOX1基因在肺、脂肪中的表达量差异极显著(P<0.01)。HMOX1基因Exon3区检测到4个SNPs位点和A、B及C 3个等位基因;通过2个品种间不同基因型比较发现,湖羊的AB型个体血红蛋白含量(hemoglobin, HGB)和红细胞压积(hematokrit, HCT)值明显低于藏绵羊的其他基因型(P<0.01),藏绵羊的AA和BB型个体的HGB和HCT值显著高于AB型(P<0.01),AC型血氧饱和度(oxygen saturation, SO2)值显著高于AA和BB型个体;而c.3775G>A位点上MM基因型个体的SO2、HGB、HCT值显著高于NN基因型(P<0.05)。由此说明AA、BB和AC基因型的c.3775G>A位点变异与血液指标存在相关性,且为G时更有利于对低氧环境的适应。因此,HMOX1基因可作为低氧适应的候选基因,为探索藏绵羊低氧适应分子机制提供基础资料。
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何建文
张伟
刘秀
李少斌
王继卿
胡江
罗玉柱
关键词 藏绵羊血红素加氧酶1基因(HMOX1)组织表达多态性血液分析    
Abstract:Heme oxygenase 1 (HMOX1) is a stress-inducible enzyme of the heme oxygenase family. HMOX1 plays an important role in the metabolic regulation of hypoxia stress in the body. Tibetan sheep (Ovis aries)have developed a unique hypoxic adaptation strategy during long-term evolution to maintain their own oxygen balance. In order to investigate the relationship between HMOX1 expression and Exon3 polymorphism and hypoxia adaptation in Tibetan sheep, look for molecular markers relating to hypoxia adaptation, this study used Tibetan sheep and Hu sheep (control group) at different altitudes, the expression of tissues and the genotype polymorphisms in exon3 of HMOX1 gene were detected by qRT-PCR and PCR-single-strand conformation polymorphism (PCR-SSCP), and the associations of different genotypes and alleles with blood parameters of sheep were analyzed. Sexuality and the correlation between different genotypes of single polymorphic loci and blood parameters of sheep were analyzed. The results showed that HMOX1 was expressed in tissues of Tibetan sheep and Hu sheep. In Tibet sheep, the gene was highly expressed in spleen (P<0.01), followed by liver, kidney and longissimus dorsi muscle (P<0.05). In Hu sheep, the gene was highly expressed in lung and fat (P<0.01), followed by liver, kidney and spleen (P<0.05). The expressions of HMOX1 gene in lung and fat tissue were significantly different between Tibetan sheep and Hu sheep (P<0.01). Four SNPs loci and 3 alleles of A, B and C were detected in the Exon3 region of HMOX1 gene. By comparing different genotypes between the two varieties, the hemoglobin (HGB) and hematokrit (HCT) values of AB Hu sheep were significantly lower than those of Tibetan sheep (P<0.01). The HGB and HCT values of BB individuals were significantly higher than those of AB (P<0.01), and the AC-type SO2 values were significantly higher than those of AA and BB individuals and c.3775G>A sites. The SO2, HGB and HCT values of MM genotypes in c.3775G>A locus were significantly higher than those of NN genotypes (P<0.05), which indicated that the C.3775G>A locus G of AA, BB and AC genotypes was more favorable for adaptation to hypoxic environment. Therefore, HMOX1 gene can be used as candidate gene.These results provide basic data for exploring the molecular mechanism of hypoxia adaptation in Tibetan sheep.
Key wordsTibetan Sheep    Heme oxygenase1 gene (HMOX1)    Tissue expression    Polymorphism    Blood analysis
收稿日期: 2018-10-28     
ZTFLH:  S826.8+3  
基金资助:甘肃省基础研究创新群体(No. 17JR5RA137)、甘肃省自然科学基金(No. 1606RJZA074)、甘肃省农业生物技术研发项目(No. GNSW-2016-24)、甘肃省国际科技合作专项(No. 18YF1WA082)和甘肃省高等学校科研项目(No. 2017B27)
通讯作者: *luoyz@gsau.edu.cn;liuxiu@gsau.edu.cn   
引用本文:   
何建文, 张伟, 刘秀, 李少斌, 王继卿, 胡江, 罗玉柱. 藏绵羊HMOX1基因表达及其多态性与低氧适应性的关联分析[J]. 农业生物技术学报, 2019, 27(3): 441-448.
HE Jian-Wen, ZHANG Wei, LIU Xiu, LI Shao-Bin, WANG Ji-Qing, HU Jiang, LUO Yu-Zhu. HMOX1 Gene Expression and Assiciation Between Its Polymorphism and Hypoxia Adaptation in Tibetan Sheep (Ovis aries). 农业生物技术学报, 2019, 27(3): 441-448.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/10.3969/j.issn.1674-7968.2019.03.007     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2019/V27/I3/441
 
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