骨多形性蛋白2cDNA在COS细胞和B16F1细胞中的表达差异

The Comparison of Expression of Bone Morphogenetic Protein 2(BMP-2)in COS Cells with That in B16F1 Cells

目的研究骨多形性蛋白2cDNA(BMP2cDNA)转染入COS细胞和B16F1细胞中的表达差异。方法BMP-2cDNA克隆在AD1-1载体中巨细胞病毒的早期启动子下游,构建了表达质粒BMP-2/AD1-1。采用脂质体法将表达质粒分别转染入COS细胞和B16F1细胞中。72 h后,分别取100μL培养上清进行Western印迹,检测所表达的BMP-2蛋白。结果Western印迹结果显示:BMP-2/Ad1-1转染的COS细胞上清液中有2条BMP-2特异表达带,大小约为39 kD和36 kD;BMP2/Ad1-1转染B16F1细胞上清液中有1条特异表达带,大小约为15 kD。结论BMP-2cDNA在COS细胞和B16F1细胞中表达产物的种类与大小均有差异。

Objective To and B16F1 cells. Methods compare the differences of expression of BMP-2 cDNA in COS cells BMP-2 cDNA was cloned into ADI-1 vector under the control of CMV promoter to construct the expression plasmid BMP-2/AD1-1. The expression vector BMP2/AD1- 1 was then transfected into COS cells and B16F1 cells respectively using cationic lipid as gene delivery vector. At 72 h post-transfection, 100 μL of culture medium was taken respectively from two transfected cells for Western blot to determine the expressed products of BMP-2. Results Two specifically expressed products, 39 and 36, of BMP-2 were detected by western blot in culture medium of the transfected COS cells. Only one specifically expressed products, 15,of BMP-2 was detected in culture medium of B16F1 cells. Conclusion COS cells express different size proteins of BMP-2, compared with B16F1 cells.