Oleosin与GUS融合蛋白基因的植物表达载体的构建及融合蛋白性质的预测

The Construction of Plant Expression Vector p^(BI-LEA-O::G121) of Oleosin and GUS Fusion Protein Gene and the Forecast of Its Properties

将油菜油脂蛋白Oleosin基因与报告基因GUS通过6个氨基酸编码的凝血酶识别位点(Gly-Val-Arg-Gly-Pro-Arg)连接起来,形成一个融合蛋白。其中编码区基因长2415bp,共编码804个氨基酸和一个终止密码子。使用蛋白质分析软件Antheprot.5.0和DNAstar对融合蛋白的理化特性,二级结构,潜在信号肽断裂位点,跨膜区进行分析表明:融合蛋白较好的保持了原来两个蛋白的理化特性和二级结构。将该融合基因克隆到中间载体pUC-121上,并用棉花LEA蛋白D-113基因启动子替换CaMV35S启动子,构建成植物表达载体pBI-LEA-O::G121。

Oleosin gene from Brassica napus and GUS gene were connected by thrombin with six amino acids , forming a fusion protein . This fusion protein gene has a coding region of 2415 base pairs encoding 804 amino acids and a terminator code A. The software of protein analysis (Antheprot.5.0 and DNAstar) was used, and results showed that the fusion protein kept physical and chemical properties as good as the original two proteins , and maintained the essential secondary structure of three proteins. By cloning the fusion gene to medium vector p^(UC-121), and replacing CaMV35S promoter by LEA protein D-113 gene promoter from cotton, the plant expression vector p^(BI-LEA-O:G121) was constructed.