水稻5.3 kb Gt1启动子克隆以及Gt1启动子引导优质大豆球蛋白Gy7基因表达载体构建

以越光水稻基因组为模板,在成功克隆5.3 kb谷蛋白Gt1基因5’上游和信号肽序列基础上,将其定向插入到pCAMBIA1300质粒中,构建了中间载体p1300-5.3 kb Gt1;再将富含赖氨酸的优质大豆球蛋白Gy7全基因序列及cDNA编码序列分别定向插入p1300-5.3 kb Gt1质粒上Gt1基因信号肽下游,再在2个载体Gy7全基因序列及cDNA编码序列下游都正向插入Gt1 3’UTR,完成由5.3 kb谷蛋白Gt1基因启动子及信号肽引导的大豆球蛋白Gy7基因2种表达载体的构建。此试验为利用转基因技术改良水稻稻米营养品质以及将水稻种子作为生物反应器等方面研究奠定了基础。

Cloning of Rice 5.3kb Gt1 Promoter and Construction of Expression Vectors with Glycinin Gy7 Gene under the Gt1 Promoter

5.3kb Gt1 gene 5'upstream sequence with Gt1 gene signal peptide was successfully cloned from rice cultivar Kohhihikari.It was inserted into the plasmid pCAMBIA1300,forming a intermediate vector,which was called p1300-5.3 kb Gt1.Then Lys-rich soybean Gy7 gene and its cDNA were inserted into p1300-5.3 kb Gt1,respectively.After that,expression vectors were constructed with insertion of Gt1 3'UTR downstream the Gy7 gene or its cDNA sequence,respectively.The experiment will lay a foundation for studies such as improving nutritional quality of rice through transgenic technology and applying bio-reactor with rice seeds.