鼠转化生长因子-β1基因的克隆及其在成骨细胞中的表达  

Cloning and expression of rat transforming growth factor beta1 cDNA in rat osteoblasts

作　　者：郑启新[1] 刘勇[1] 杜靖远[1] 屈伸[2] 曾晖[1] 郭晓东[1] 

机构地区：[1]同济医科大学附属协和医院骨科,武汉430022 [2]同济医科大学基础医学院生化教研室

出　　处：《中华实验外科杂志》2000年第4期344-345,共2页Chinese Journal of Experimental Surgery

摘　　要：目的　克隆鼠转化生长因子 β1(TGF β1)基因并研究其在转染的成骨细胞中表达情况。方法　采用逆转录 聚合酶链式反应 (RT PCR)可从大鼠淋巴细胞中扩增出 1.2Kb特异性片段 ,经酶切鉴定证实为鼠TGF β1cDNA。将此片段插入 5 .4Kb的 pcDNA表达载体 ,构建得到 6 .6Kb的pcDNA TGF β1重组质粒。应用脂质体介导的基因转移技术将TGF β1表达载体导入鼠成骨细胞 ,48h后用SABC法进行瞬时表达的检测。结果　 48h后转染的成骨细胞具有明显鼠TGF β1基因的表达产物。结论　将构建的TGF β1表达载体转入鼠成骨细胞 ,48h后可获得TGF β1的高效表达。Objective To clone rat transforming growth factor (TGF) beta1 cDNA, and to investigate TGF beta1 gene expression in transfected osteoblasts. Methods One amplificaiton fragment (1.2 Kb) was obtained from the rat lymphocytes by RT-PCR method and cloned into expression vector pcDNA (5.4 Kb), which was named pcDNA-TGF beta1 (6.6Kb). The cloned gene was confirmed to code for rat TGF beta1 by restriction enzyme analysis. Rat osteoblasts were transfected with pcDNA-TGF beta1 plasmid by lipofectamine mediated gene transfer. The transient expression was detected by using SABC method 48 h later.Results Rate TGF beta1 gene expression could be detected 48 h later in the transfected osteoblasts. Conclusion After rat TGF beta1 was cloned into expression vector pcDNA, TGF beta1 gene could be expressed effectively in transfected osteoblasts by pcDNA-TGF beta1 plasmid 48 h later.

关 键 词：转化生长因子-Β1 分子克隆 基因表达 成骨细胞 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]