SQ RT-PCR检测不同品种猪脂肪组织SOCS-3基因表达

mRNA Expression of SOCS-3 Gene in Adipose Tissue of Different Swine Breeds

取4月龄八眉猪和大白猪背部皮下脂肪组织,提取总RNA,根据大鼠细胞因子信号转导抑制因子-3基因(Suppressor Of Cytokine Signaling-3,SOCS-3)设计并合成引物,以猪-βactin基因作为内参,优化反应条件和体系,半定量(Semi Quantitative,SQ)RT-PCR单管扩增猪SOCS-3基因,经琼脂糖凝胶电泳分离后,Dolphin-DOC凝胶图像分析软件分析,测定各条带的光密度值,检测八眉猪和大白猪脂肪组织中SOCS-3基因mRNA的表达差异。结果表明:八眉猪脂肪组织SOCS-3 mRNA的表达丰度极显著高于大白猪(P<0.01)。这种差异可能是2种经济类型猪脂肪沉积能力不同的主要原因之一。

Various economics pig breeds show remarkable difference on their fat deposition ability. The objective of this study is to determine their genetic difference by determining their gene expression level during the fat deposition growth in Bamei pigs and Largewhite pigs. A total RNA was extracted from dorsal subcutaneous adipose tissue at 4 months old of Bamei and Largewhite pig breeds, respectively. The first-strand cDNA was synthesized by reverse transcriptase using random hexamer primer. A pair of primers were designed based on the sequence of rat Suppressor of Cytokine Signaling 3 gene （SOCS-3）. The single tube Semi Quantitative Reverse Transeription-Polymerase Chain Reaction （SQ RT-PCR） was applied to determine their expression level of SOCS 3 mRNA in adipose tissue between Bamei pig and Largewhite pig. The β-actin gene was selected as internal control. After amplication, agarose gel electrophoresis was performed and various expression bands and their Optical Density （OD） value were analyzed by Dolphin-DOC software. The result showed that the expression level of SOCS-3 mRNA in dorsal subcutaneous adipose tissue in Bamei pig was significantly higher than that of Largewhite pigs （P〈 0.01）. This finding suggests that the difference of SOCS-3 gene expression level in adipose tissue may be one of the main reasons which result in the different fat deposition ability between different pig breeds.