目的 构建脂肪酸结合蛋白3(FABP3)基因的荧光表达载体FABP3-pEGFP-N2,并应用绿荧光融合蛋白技术检测FABP3蛋白的亚细胞定位.方法 RT-PCR法获得FABP3基因编码框,克隆到pEGFP-N2载体中,利用脂质体将构建的表达载体转染P19细胞后,荧光显微镜和激光共聚焦显微镜观察细胞内FABP3蛋白定位.结果 成功克隆FABP3基因片段,并将其重组到pEGFP-N2载体中.荧光显微镜和激光共聚焦显微镜显示,FABP3蛋白主要分布于P19细胞的胞浆和胞核中.结论 成功构建了FABP3-pEGFP-N2真核表达载体,FABP3在P19细胞的胞浆和胞核中呈弥漫分布特点.%Objective To construct the recombinant fluorescence expression vector pEGFP-N2 containing fat acid binding protein 3 (FABP3) gene and investigate the sub-cellular localization of FABP3 protein by green fluorescence fusion protein technology. Methods The open reading frame sequence of FABP3 was amplified by RT-PCR and then cloned into pEGFP-N2 piasmid After the recombinant expression vector FABP3-pEGFP-N2 was transfected into P19 cells, the localization of FABP3 protein was observed by fluorescence microscope and laser confocal microscope. Results The FABP3 gene fragment was successfully cloned and recombined into pEGFP-N2 piasmid. The fluorescence microscope and laser confocal microscope showed that FABP3 protein was mainly localized in the cytoplasm and nucleus of PI 9 cells. Conclusion The eukaryotic expression vetor FABP3-pEGFP-N2 has been successfully constructed, and FABP3 is widely expressed in the cytoplasm and nucleus of PI 9 cells.
展开▼
