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 FABP3-pEGFP-N2 expression vector of FABP3 gene and detect the subcellular localization of FABP3 protein by green fluorescence fusion protein technology. Methods The coding sequence of FABP3 gene was obtained by RT-PCR and cloned into pEGFP-N2 vector. The constructed vector was transfected into P19 cells by lipofectamine. The localization of FABP3 protein was observed by fluorescence microscope and confocal laser scanning microscopy. Results FABP3 gene fragment was successfully cloned and recombined into pEGFP-N2 vector. Fluorescence microscopy and confocal laser scanning microscopy showed that FABP3 protein mainly distributed in the cytoplasm and nucleus of P19 cells. Conclusion The eukaryotic expression vector FABP3-pEGFP-N2 was successfully constructed. FABP3 was diffusely distributed in the cytoplasm and nucleus of P19 cells.