猪A-FABP基因真核表达载体的构建及其组织表达

本实验以长白猪背最长肌为实验材料,进行组织RNA的提取及A-FABP基因的克隆,构建p EGFPN1-A-FABP真核表达载体,并通过脂质体转染成纤维细胞,48 h观察荧光表达。同时应用荧光定量PCR法检测猪7种不同组织(心脏、肝脏、脾脏、肺脏、肾脏、背最长肌、腿肌)中A-FABP基因的差异表达。结果表明:成功构建p EGFP-N1-A-FABP融合表达载体,并在细胞中表达绿色荧光蛋白;A-FABP基因在7种组织中均有表达,其中背最长肌和腿肌中A-FABP基因的表达量最高,与其他组织相比差异极显著(P0.01),而脾脏和肾脏中的表达量相对于心脏、肝脏和肺脏差异显著(P0.05),而A-FABP基因表达量在心脏、肝脏和肺脏中差异不显著(P0.05)。表明该基因能够在真核载体和细胞中表达,并且在猪不同组织的表达具有差异性。

Constructing of Eukaryotic Expressing Vector and Tissues Expression of A-FABP Gene in Pig

The longissimus dorsi muscle of Landrace pig was as experimental material, RNA was extracted and A-FABPgene was cloned, the eukaryotic expression vector of p EGFP-N1-A-FABP was constructed and transfected to fibroblasts by liposome, the fluorescence expression was detected after 48 h. Then the differential expression of A-FABP gene in different tissues(heart, liver, spleen, lung, kidney, longissimus dorsi muscle and leg muscle) was detected by Q-PCR. Results showed that the p EGFP-N1-A-FABP fusion expression vector was successfully constructed, the fluorescence expression was successfully expressed in fibroblasts. The expression of A-FABP gene in 7 different tissues was different. The gene expression in longissimus dorsi muscle and leg muscle were the highest compared with others, they had extremely significant difference(P<0.01), the expression in spleen and kidney were significantly different from that of heart, liver and lung(P<0.05), while the A-FABP gene was not significantly different in heart, liver and lung. Results indicated that A-FABP gene was expressed in eukaryotic vector and cells, and its expression had difference in different tissues of pigs.