从江香猪LMF1基因的克隆、亚细胞定位及组织表达研究

Cloning,Subcellular Localization and Tissue Expression of LMF1 Gene in Congjiang Xiang Pig

为研究脂肪酶成熟因子1(LMF1)基因在从江香猪不同组织的表达情况及在真核细胞中的亚细胞定位情况,采用RT-PCR,Real-Time PCR,PSORT II Prediction软件结合荧光共定位的方法,克隆了从江香猪LMF1基因cDNA,构建了pEGFP-C1-LMF1真核表达载体,分析了LMF1基因在不同组织的表达差异和亚细胞定位情况.结果表明,从江香猪LMF1基因与GenBank上提交的其他猪种的同源性达到了100%;LMF1基因在脂肪中表达量最高,心中最低,且脂肪和小肠与其他7个组织相比,表达量均存在统计学意义(p<0.01);荧光共定位结果表明,LMF1基因主要集中在细胞质中表达.相关结果对后续研究LMF1蛋白与其互作蛋白在脂质代谢的作用机理奠定的基础.

In order to study the expression of lipase maturation factor 1（LMF1 gene）in different tissues of Congjing Xiang pig and its subcelluar localization,RT-PCR（real-time PCR）,PSORT II Prediction and fluorescent co-location were used to clone the cDNA of LMF1 gene,construct the recombinant eukaryotic vector of pEGFP-C1-LMF1,and analyze the difference of its expression in various tissues and subcelluar localization,so as to provide a foundation for researching the mechanism of LMF1 protein and its interaction proteins in lipid metabolism.The results showed that the cDNA region of LMF1 gene of Congjiang Xiang pig and that of other pig breeds in GenBank submission shared a 100% homology.LMF1 gene had the highest expression in the fat and the lowest in the heart,and its expression in the fat and the small intestine was highly different from that in the other 7 tissues（p0.01）.Fluorescent co-location demonstrated that LMF1 gene was mainly expressed in the cytoplasm.