本试验采用单核苷酸多态性(single nucleotide polymorphism,SNP)直接测序法检测猪锚蛋白重复序列和SOCS盒蛋白基因4(ankyrin repeat and SOCS box containing protein 4,Asb4)与生长因子受体结合蛋白基因10(growth factor recep-tor-bound protein 10,Grb10)在不同组织器官中的印迹状态。首先,克隆得到了1350bp的Asb4基因cDNA序列及1811bp的Grb10基因cDNA序列,然后进行了SNP直接测序法检测。结果发现,Asb4在1月龄仔猪所有检测组织器官中均为双等位基因表达,而Grb10在1月龄仔猪的舌、肾脏、胃、小肠和脑中为父源等位基因表达,在其他组织器官中为双等位基因表达。实时定量PCR结果表明,Grb10在10种组织器官中的表达量存在显著性差异(P〈0.05),其中在肺脏组织中的表达量最高,在小肠和脑中表达量最低。上述结果表明,Grb10可能是猪父源表达的印迹基因,而Asb4则属于猪非印迹基因。
In this study, by using a single nucleotide polymorphism-based method, 1350 bp cDNA sequence of porcine ankyrin repeat and SOCS box containing protein 4 (Asb4) and 1811 bp cDNA sequence of porcine growth factor receptor-bound protein 10 (Grbl0) was obtained. Then we identified their imprinting status in ten tissues. Imprinting analysis showed that both alleles of Asb4 were expressed in all the tested tissues. Grbl0 was paternally expressed in tongue, kidney, stomach, in- testine and brain, while biallelically expressed in the others. We also used quantitative Real-time PCR to assess the expression of Grbl0 in these tissues. The expression level of Grbl0 was significantly different among tissues, which was the highest in lung and lowest in both small intestine and brain. These results primarily indicated that Grbl0 probably was a porcine paternal- ly expressed gene, but Asb4 was not imprinted in swine.