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目的构建pEGFP-C1-PEX真核表达载体并转染大鼠骨髓间质干细胞(MSCs),观察其在MSCs内的表达。方法用RT-PCR法从大鼠C6胶质瘤细胞中扩增出带有XhoI、BamHI酶切位点的PEX基因片段,将PEX基因片段克隆到增强型绿色荧光蛋白(EGFP)基因真核表达载体pEG-FP-C1上,通过脂质体将pEGFP-C1-PEX转染入大鼠MSCs。结果pEGFP-C1-PEX真核表达载体构建成功,荧光显微镜下观察,pEGFP-C1-PEX表达载体转染到MSCs24h后可见PEX融合蛋白表达。结论pEGFP-C1-PEX真核表达载体能够在大鼠MSCs中表达PEX融合蛋白。
Objective To construct pEGFP-C1-PEX eukaryotic expression vector and transfect rat bone marrow mesenchymal stem cells (MSCs) to observe its expression in MSCs. Methods PEX gene fragment with XhoI and BamHI restriction sites was amplified from rat C6 glioma cells by RT-PCR. The PEX gene fragment was cloned into eukaryotic expression of enhanced green fluorescent protein (EGFP) gene On the pEG-FP-C1 vector, pEGFP-C1-PEX was transfected into rat MSCs by liposome. Results The eukaryotic expression vector pEGFP-C1-PEX was constructed successfully. The expression of PEX fusion protein was observed under fluorescence microscope 24 h after transfection with pEGFP-C1-PEX expression vector. Conclusion The pEGFP-C1-PEX eukaryotic expression vector can express PEX fusion protein in rat MSCs.