稳定表达人骨形成蛋白2成纤维细胞的建立与鉴定

Establishment and identification of stably-expression human hone morphogenetic protein 2 fihroblasts

目的：探讨BMP对细胞分化的调节作用，为BMP基因疗法的建立提供依据．方法：构建了含有人BMP2开放阅读框（1．2kb）的真核表达载体PBK－B2，利用脂质体转染方法将其导入NIH3T3细胞，通过G－418筛选获得阳性细胞克隆．细胞原位杂交和免疫组化检测BMP2基因的稳定转染及表达情况．结果：转染细胞内有BMP2mRNA的转录及其蛋白的表达．结论：人BMP2基因在NIH3T3细胞中得到稳定有效的表达．

AIM: To elucidate the relationship between BMPand cell differentiation, and to provide basis for BMP genetherapy. METHODS: Eukaryonic expression vector (pBKB2 ) containing human BMP2 open reading frame (1. 2 kb)was constructed and transduced into NIH3T3 cells by usinglipofectamine. The positive cell clones were selected with G-418. The stable transfection and expression of BMP2 in theNIH3T3 cells were determined by in situ hybridization and immunohistochemical methods.RESULTS: BMP2 mRNA wastranscripted and expressed in the trans feeted cells. CONCLUSION: Human BMP2 gene is stably and effectively expressedin the NIH3T3 cells.