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目的 探讨外源性hBMP2 基因在成纤维细胞获得稳定表达的可行性。 方法 将hBMP2 的cDNA连入真核表达载体pcDNA3 ,形成重组真核表达载体pcDNA3 hBMP2 ,在脂质体介导下 ,将其导入小鼠成纤维细胞株 (NIH3T3)。通过G418筛选获得阳性克隆 ,并继续培养 4周 ,然后用原位杂交和免疫组织化学方法检测BMP2 基因在成纤维细胞NIH3T3内的稳定表达情况。 结果 经原位杂交和免疫组织化学证实 ,转染pcDNA3 hBMP2 后的成纤维细胞NIH3T3内有大量hBMP2 mRNA的转录和蛋白的表达。 结论 在脂质体介导下 ,hBMP2 基因能够导入成纤维细胞内并在其内获得稳定表达
Objective To investigate the feasibility of stable expression of exogenous hBMP2 gene in fibroblasts. Methods The cDNA of hBMP2 was inserted into eukaryotic expression vector pcDNA3 to form recombinant eukaryotic expression vector pcDNA3 hBMP2. The recombinant plasmid was introduced into NIH3T3 mouse fibroblast cell line mediated by liposome. Positive clones were screened by G418 and cultured for 4 weeks. Then, the stable expression of BMP2 gene in NIH3T3 fibroblasts was detected by in situ hybridization and immunohistochemistry. Results The in situ hybridization and immunohistochemistry confirmed that a large number of hBMP2 mRNA transcription and protein expression were found in NIH3T3 fibroblasts transfected with pcDNA3 hBMP2. Conclusion The hBMP2 gene can be introduced into fibroblasts under the liposome-mediated and stable expression