关键词: 软骨细胞, 骨形成蛋白-7, 真核表达载体

Abstract: Objective To construct the eukaryotic expression vector of pcDNA3.1-BMP7 and transfect it into the articular chondrocytes of rabbit cultured in vitro in order to study the best condition of liposome and plasmid DNA. Methods Expression vector of pcDNA3.1-BMP7 was constructed and was digested by two enzymes and sequenced. pcDNA3.1-BMP7 was transfected into the articular chondrocytes of rabbit mediated with liposome and screened by G₄₁₈.The best rates of liposome and G₄₁₈, liposome and pcDNA3.1-B MP7 were identified. In situ hybridization, PCR and Western blotting were used to detect the expression of BMP7.Results The BMP7 gene of pcDNA3.1-BMP7 was same as the BMP7 gene reported.The best concentration was 10 μL liposome and 4 μg pcDNA3.1-BMP7 in 3 mL IMDM medium including 400 mg·L^-1 G₄₁₈. Conclusion Madiated with liposome reagent, the articular cartilages of rabbit are successfully transfected with pcDNA3.1-BMP7 and BMP7 express stably in the articular cartilages.

Key words: chondrocytes, bone morphogenetic protein-7, eukaryotic expression vector