藏猪和大约克猪CKMT2基因多态性分析及其组织表达模式

doi:10.16431/j.cnki.1671-7236.2020.10.030

摘要/Abstract

摘要： 为探究藏猪和大约克猪肌节线粒体肌酸激酶2（creatine kinase，mitochondrial 2，CKMT2）基因多态性位点及其在各组织中的表达差异，试验采用Sanger测序法对藏猪（50头）和大约克猪（60头）CKMT2基因起始密码子上游1 000 bp区域进行单核苷酸多态性（single nucleotide polymorphism，SNP）筛选与基因分型；180日龄藏猪和大约克猪（各10头）屠宰后分别采集心脏、背脂和背最长肌组织，采用实时荧光定量PCR技术检测CKMT2基因在各组织中的表达水平。结果显示，在CKMT2基因起始密码子上游1 000 bp区域，筛选到5个SNPs位点：G-357A、T-469C、G-649T、A-784G和A-953G，且藏猪与大约克猪等位基因频率差异明显。转录因子预测发现，这些SNPs位点与骨骼肌生长发育、肌肉能量代谢、抗缺氧有关。藏猪CKMT2基因在背最长肌和心脏组织中的mRNA表达量极显著高于大约克猪（P<0.01），在背脂中的mRNA表达量极显著低于大约克猪（P<0.01），推测CKMT2基因具有正向调控抗低氧和肌肉组织生成、负向调控脂肪生成的作用，该分析结果与藏猪和大约克猪的品种特性相吻合。本试验结果为进一步探索猪生长发育、脂肪沉积和低氧适应性提供了一定的新思路。

关键词: 藏猪; 大约克猪; CKMT2基因; 多态性; mRNA表达

Abstract: In order to explore the polymorphism site and difference of expression level of creatine kinase,mitochondrial 2 (CKMT2) gene in muscle segments of Tibetan and Yorkshire pigs,in this experiment,Sanger sequencing method was used to screen and genotype the single nucleotide polymorphism (SNPs) in the 1 000 bp region upstream of the initiation codon of CKMT2 gene in 50 Tibetan and 60 Yorkshire pigs.The expression of CKMT2 gene was detected by Real-time quantitative PCR in heart,back fat and longissimus dorsi muscle of 180-day-old of 10 Tibetan and 10 Yorkshire pigs.The results showed that 5 SNPs (G-357A,T-469C,G-649T,A-784G and A-953G) were found in the upstream 1 000 bp region of the initiation codon of CKMT2 gene,the allele frequency of Tibetan pigs at all sites were significantly different from those of Yorkshire pigs.These SNPs were related to skeletal muscle growth and development,muscle energy metabolism and anti-hypoxia.The mRNA expression of CKMT2 gene in longissimus dorsi muscle and heart of Tibetan pigs were extremely significantly higher than Yorkshire pigs (P<0.01),and the mRNA expression in back fat of Tibetan pigs was extremely significantly lower than Yorkshire pig (P<0.01),which indicated that CKMT2 gene could positively regulate anti-hypoxia and muscle tissue production,and negatively regulate fat production,the analysis results were consistent with the breed characteristics of Tibetan and Yorkshire pigs.This experiment results provided some new ideas for further exploration of pig growth and development,fat deposition and hypoxia adaptability.

Key words: Tibetan pigs; Yorkshire pigs; CKMT2 gene; polymorphism; mRNA expression

中图分类号:

S813.3

王珍梅, 段梦琪, 陆锋辉, 王士欣, 强久卓玛, 谭占坤, 张健, 商鹏. 藏猪和大约克猪CKMT2基因多态性分析及其组织表达模式[J]. 中国畜牧兽医, 2020, 47(10): 3305-3313.

WANG Zhenmei, DUAN Mengqi, LU Fenghui, WANG Shixin, QIANGJIUZHUOMA, TAN Zhankun, ZHANG Jian, SHANG Peng. Polymorphism and Tissue Expression Patterns of CKMT2 Gene in Tibetan and Yorkshire Pigs[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(10): 3305-3313.

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