花生白藜芦醇合酶基因转化单子叶植物表达载体的构建

Construction of the Monocotyledon Expression Vector for the Transforming of Resveratrol Synthase Gene

构建了花生白藜芦醇合酶基因(RS)转化单子叶植物的表达载体,该表达载体含有ub i启动子和内含子,能启动该基因在单子叶植物中高效地表达.通过PCR反应扩增出目的片段,连接到克隆载体Pub i35s上,切下含ub i和RS约3 000 bp的片段连接到植物表达载体pCAM B IA-1 380上.经PCR和酶切检测,结果与预期相同,经测序确定插入片段读码框正确.该表达载体可用于单子叶植物高效的表达.

A monocotyledon expression vector was constructed for transforming Resveratrol Synthase （RS） gene. This vector contains ubi promoter and intron which can drive high expression of the gene in monocotyledon . PCR was used to amplify the RS genc. A new cloning vector was then constructed by inserting the amplified RS gene into Pubi35s. The monocotyledon expression vector was constructed by inserting the ubi promoter and RS gene （about 3 000 bp） into pCAMBIA-1 380, which cutted from the new cloning vector above. PCR and restriction enzyme digesting analysis showed that the inserting DNA fragment was the size of expecting. Furthermore , the sequencing showed that the insert DNA fragment was not out of frame. The expression vector can be used for the high expression in monocotyledon.