pcDNA3.1-BMP-2真核表达载体构建及转染兔骨髓基质细胞的实验研究

Construction of mammal expression plasmid pcDNA3. 1-BMP-2 and transfection of bone marrow stromal cells

目的 构建人骨形态发生蛋白2(BMP-2)真核表达载体,鉴定并转染兔骨髓基质细胞,为转基因治疗骨缺失疾病提供实验基础。方法 双酶切克隆载体将BMP-2目的基因与真核表达载体pcDNA3.1连接,免疫组化及Western blotting检测转染BMP-2基因骨髓基质细胞的表达效果。结果 成功构建BMP-2真核表达载体,转染骨髓基质细胞后,有BMP-2表达。结论 骨髓基质细胞经基因转染后可以表达BMP-2,为进一步实验研究提供基础。

Objective To construct mammal expression plasmid vector containing human BMP-2 for the study of gene therapy of bone defect disease. Methods Clone plasmid DNA pspBMP-2 was digested by Xba I and Sal I, recombined with plasmid pcDNA3.1, and the plasmid was transfected into rabbit bone marrow stromal cells. The expression of bone morphogenetic protein-2 was detected by immunochemistry and Western blotting. Results After BMP-2 expression plasmid was constracted, bone morphogenetic protein-2 was detected in bone marrow stromal cells. Conclusion Bone marrow stromal cells can express bone morphogenetic protein-2 after transfec-tion. This method provides a basis for studying BMP-2.