猪IGF-1R胞外域编码区2种单倍型对基因表达的影响

Effects of Two Haplotypes in Extracellular Coding Region of Porcine IGF-1R on Gene Expression

试验从筛查到的大型猪和小型猪胰岛素样生长因子1受体(IGF-1R)基因胞外域编码区上的4个同义突变形成的2种单倍型入手,分析不同猪种IGF-1R的2种单倍型是否影响其转录和翻译。通过构建包含IGF-1R 2种单倍型的表达载体(大型猪组命名为LP组,小型猪组命名为BM组),利用qRT-PCR和Western blot,在转录和翻译水平上分析大型猪和小型猪IGF-1R的2种单倍型对基因表达的影响。结果表明:LP组的IGF-1R mRNA表达量显著高于BM组(P<0.01);进一步检测发现,与LP组相比,BM组的IGF-1R蛋白质表达量显著降低(P<0.05)。大型猪和小型猪IGF-1R胞外域编码区2种单倍型在转录和翻译水平影响基因表达,进而可能调控动物的生长发育,为进一步研究小型猪形成机制提供数据。

Insulin-like growth factor 1 receptor(IGF-1R)plays an important role in regulating animal growth and development.Multiple single nucleotide polymorphisms(SNPs)were found in IGF-1R gene between large pigs and miniature pigs.Starting from the two haplotypes formed by four synonym mutations in the extracellular coding region of IGF-1R gene of screened BaMa miniature and Yorkshire pigs,we analyzed whether the two haplotypes of IGF-1R in different pig breeds affected its transcription and translation.By constructing expression vectors containing the two haplotypes of IGF-1R(LP group for large pig group and BM group for miniature pig group),the effects of the two haplotypes of IGF-1R on gene expression in large pig and miniature pig groups were analyzed at the transcription and translation levels by qRT-PCR and Western blot.The results showed that the mRNA expression of IGF-1R in LP group was significantly higher than that in BM group(P<0.01).Further testing showed that compared with the LP group,the expression of IGF-1R protein was significantly decreased in the BM group(P<0.05).In conclusion,the two haplotypes of IGF-1R extracellular domain coding region in large and miniature pigs affect gene expression at the transcription and translation levels,which may regulate the growth and development of animals,and provide data for further study on the formation mechanism of miniature pigs.