油体蛋白与人成骨蛋白-1基因融合植物表达载体的构建及融合蛋白性质预测

Construction of Plant Expression Vector of Oleosin and Bmp7 Fusion Gene and Prediction Properties of Recombinant Protein

克隆大豆油体蛋白特异启动子及油体蛋白基因的全长序列,利用连接PCR的方法,通过一个肠肽酶位点将人成骨蛋白成熟肽编码序列连接到油体蛋白基因3’端并克隆到表达载体pCAMB IA1302中,构建"油体蛋白-肠肽酶-人成骨蛋白"植物表达载体。通过酶切、PCR和测序鉴定,证明表达载体构建成功,将其命名为pCAM-oleo-Bmp7。通过蛋白分析软件DNAstar、S ignalP3.0和ProtComp v8.0等软件对重组蛋白的理化性质、二级结构、信号肽和细胞定位进行了分析和预测,证明融合蛋白很好的保留了原来两个蛋白的二级结构和理化性质。

The oleosin gene and promoter was cloned from soybean.By ligation PCR methods,oleosin gene and Bmp7 gene was connected by the enterokinase gene and the fusion gene was constrcuted in pCAMBIA1302.After proven by restriction enzyme digestion and PCR,the expression vector was named pCAM-oleo-Bmp7.The signal peptide、sub-cellular localization and secondary structure of the fusion protein were predicted.The results showed that the fusion protein kept the physical and chemical properties of the original proteins＇ .