摘要
目的构建hBMP2真核表达载体pcDNA3-hBMP2,将其转染兔骨髓基质细胞(Marrow Stromal Cells,MSCs)并检测其表达效率。方法将hBMP2的cDNA构建于真核表达载体pcDNA3,形成重组真核表达载体pcDNA3-hBMP2,酶切鉴定后体外转染培养状态下的兔骨髓基质细胞,用原位杂交和免疫组织化学方法鉴定表达情况并用计算机图像分析系统测定其表达效率。结果pcDNA3-hBMP2载体酶切鉴定与预期片段相符,表明成功构建了pcDNA3-hBMP2转基因载体。用该载体转染兔骨髓基质细胞后获得了瞬时表达和稳定表达。结论载体pcDNA3-hBMP2可以在骨髓基质细胞中表达,为下一步将其用于转基因骨组织工程研究奠定基础。
Objective To construct human bone morphogenetic protein-2 （ hBMP-2 ） expressing eukaryotic vector （ pcDNA3-hBMP2 ） and examine its expression in rabbit bone marrow stremal cells （MSCs） . Methods pcDNA3-hBMP2 vector was constructed using gene clone and recombination technique. After being confirmed by enzyme cut, pcDNA3-hBMP2 was transfected into cultured rabbit MSCs in vitro with the help of liposome reagent FuGENE6. The expression of hBMP-2 was determined by in situ hybridization and immunohistochemical analysis. Results The pcDNA3-hBMP2 vector was constructed successfully. The rabbit MSCs transfected by pcDNA3-hBMP2 showed transient and stable expression of BMP2. Conclusion pcDNA3-hBMP2 can express hBMP2 in bone marrow stromal cells.
出处
《中国实验动物学报》
CAS
CSCD
2006年第1期44-46,T0005,共4页
Acta Laboratorium Animalis Scientia Sinica
