迪庆藏猪ADFP基因多态性及其组织表达特征

The polymorphism and its expression characteristics in the tissues of ADFP gene in Diqing Tibetan pig

  • 摘要: 【目的】从分子遗传学角度分析不同基因型迪庆藏猪群体不同组织中ADFP基因的表达差异,为迪庆藏猪的选育工作提供参考依据。【方法】利用PCR扩增测序方法对113头迪庆藏猪ADFP基因进行扩增测序,利用DNA-STAR 5.0对序列进行比对分析及多态性位点(SNPs)检测;同时利用实时荧光定量PCR方法对迪庆藏猪不同组织中ADFP基因相对表达量进行检测,并运用SPSS 20.0分析ADFP基因多态性对迪庆藏猪群体不同组织中ADFP基因表达水平的影响。【结果】在迪庆藏猪ADFP基因外显子6上发现3个SNPs,分别为SNP1(C→T)、SNP2(C→T)和SNP3(T→ C),其中SNP1和SNP2位点属于低度多态(PIC/He<0.25),SNP3位点属于中度多态(0.25He<0.5)。迪庆藏猪不同组织中ADFP基因相对表达量存在显著差异(P<0.05,下同),皮下脂肪中相对表达量最高,其次为肺脏、肝脏、脾脏、肾脏、腿肌和背肌,心脏中相对表达量最少;另外,ADFP基因SNPs不同基因型迪庆藏猪群体中各组织的相对表达量也存在显著差异,尤其在SNP2位点杂合基因型群体肾脏中相对表达量显著高于纯合基因型群体肾脏中相对表达量,表明ADFP基因SNPs变异对迪庆藏猪组织中ADFP基因表达具有影响。【结论】ADFP基因SNPs变异对迪庆藏猪组织中ADFP基因会产生差异表达,进而影响机体脂肪沉积,ADFP基因可作为迪庆藏猪肌内脂肪沉积的候选基因。

     

    Abstract: 【Objective】This study aimed to analyze the expression difference of ADFP gene in different tissues of Diqing Tibetan pig with different genotypes from the perspective of molecular genetics, so as to provide a reference for breeding of Diqing Tibetan pig.【Method】In this study, the ADFP gene of 113 Diqing Tibetan pigs were amplified using PCR and sequenced, and the sequence alignment and the polymorphic sites(SNPs) were detected with DNASTAR 5.0. Meanwhile, the real-time fluorescence quantitative PCR(qPCR) was used to detect the relative expression of the ADFP gene in tissues of Diqing Tibetan pigs, and the SPSS 20.0 was used to analyze the effect of ADFP gene polymorphism on its expression in the tissues of Diqing Tibetan pigs.【Result】Three SNP1(C→T), SNP2(C→T) and SNP3(T→C) were found on exon 6 of ADFP gene in Diqing Tibetan pig, and SNP1 and SNP2 were low polymorphic(PIC/He<0.25) and SNP3 was moderate polymorphic(0.25<PIC/He<0.5). There were significant differences in the relative expression levels of ADFP gene in different tissues of Diqing Tibetan pigs(P<0.05, the same below). Subcutaneous fat had the highest relative expression level, followed by lung, liver, spleen, kidney, leg muscle and back muscle, heart had the lowest expression level. In addition, there were also significant differences in the relative expression levels of ADFP gene polymorphisms among different genotypes in the Diqing Tibetan pig. Especially, the relative expression of the gene in the heterozygous genotype groups at SNP2 was significantly higher than that in the homozygous genotype groups, the results showed that the polymorphisms of ADFP gene had an effect on the expression of ADFP gene in tissues of Diqing Tibetan pig.【Conclusion】The difference of ADFP gene polymorphism sites can cause differential expression of ADFP gene in tissues of Diqing Tibetan pig, and then affect fat deposition in the body, ADFP gene can be used as a candidate gene for fat deposition in Diqing Tibetan pig。

     

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