猪红细胞生成素受体基因突变与血常规性状关联分析

Association of SNPs in Erythropoietin Receptor Gene with Blood Parameters

本研究旨在探索红细胞生成素受体基因(EPOR)影响红细胞相关性状的分子机理。对EPOR基因第1内含子和第4内含子突变位点g.705G>T和g.2373C>T,以飞行时间质谱进行个体基因型分型,在构建的大白猪×民猪F2代资源群体内开展其与6种血常规性状(红细胞压积、血红蛋白、平均红细胞血红蛋白量、平均红细胞体积、红细胞计数及红细胞分布宽度)的关联分析。结果表明,第1内含子突变位点g.705G>T未发现与血常规性状显著关联;第4内含子突变位点g.2373C>T群体内只发现2种基因型,其中CT基因型个体的红细胞压积显著高于CC基因型个体(P<0.05),但并未发现与其他性状显著关联。结果显示,EPOR基因突变可显著影响红细胞压积,说明该基因可作为影响红细胞压积的主效候选基因开展深入研究。

The aim of this study was to reprogram fibroblasts cell by in vitro transcribed mRNA cocktail of Oct4, Sox2 and SV40 T-antigen, and provide a safe, nonintegrating strategy for somatic cell reprogramming. The recombinated mRNA transcription vectors including the above three induced facotrs genes were constructed and transcripted, respectively, then 5′UTR and 3′ UTR of β-globin gene were used for stabilizing in vitro transcribed mRNA following in vitro capping and poly（A） tailing. After mRNA transfection of 293 and IMR90 cells, immunocytochemistry, immunoflurescence and Real-Time PCR methods were used to detect gene expression, protein location and endogenous gene expression related to pluripotence. The results showed that target gene expressions could be detected in 293 and IMR90 cells and all expressed protein were localized properly in the nucleus. The specific expression of Oct4 and Nanog were increased in transfection cells. Endogenous Nanog expression was induced by mRNA cock-tail transfection. These findings indicate that mRNA of Oct4, Sox2 and SV40 T in vitro can coorperate to initiate cellular reprogramming.