BMP-4基因在成纤维细胞内的表达

Expression of BMP-4 Gene in Fibroblasts

目的 :构建mBMP 4真核表达载体并转染成纤维细胞 ,观测外源基因在靶细胞内的表达及对细胞表现型的影响。方法 :利用基因重组技术构建mBMP 4基因真核表达载体 pCR 3 .1uni mBMP 4;利用基因转染技术体外转染mBMP 4基因至成纤维细胞NIH 3T3 ,转染的细胞用G418筛选 ,利用斑点杂交和免疫组化检测转染 4周后外源基因表达的情况。结果 :琼脂糖电泳证明mBMP 4真核表达载体 pCR 3 .1uni mBMP 4构建成功。细胞转染后第 4周 ,斑点杂交和免疫组化均显示为阳性 ,阳性细胞碱性磷酸酶表达升高。结论 :外源BMP 4基因可在成纤维细胞内获得表达并可影响靶细胞的表现型。

Objective: To construct the eukaryonic expression vector of mBMP 4,transfes the eukaryonic expression vector pCR 3.1uni mBMP 4 into the fibroblast and observe the expression of the transferred gene in the cells.Method:Using the gene recombine technology to construct the eukaryonic expression vector pCR 3.1uni mBMP 4.With the help of lipofectamine and using the gene transferring technology,we transferred the expression vector into the fibroblast NIH3T3.The transferred cells were filtered by G418.The exprexssion of mBMP 4 was evaluated by stain hybrization and immuno histochemistry technology 4 weeks after the transfer.Result:Agarose electrophoresis proved the success of the construction of the eukaryonic expression vector of mBMP 4.Through stain hybrization and immuno histochemistry,we proved that the transferred cells had expressed mBMP 4 at mRNA and protein levels after 4 weeks of filteration with G418.The expression of ALP of the positive cells had also increased.Conclusion:Heterogenous mBMP 4 gene could be expressed in the target cell fibroblast.The expression of the gene could also affect the phenotype of the target cells. Key words Bone morphogenetic protein Fibroblast Gene therapymptoms of bowe l obstruction usually occurred about sev en days after operation,mostly caused by extensive inflammations in abdomen;②Abd ominal distension more prominent than ab dominal pain,and rare intestinal ischemi a;③Non-operative treatment should be the first choice for EPISBO.