苜蓿花叶病毒复制酶基因全长cDNA植物表达载体的构建和对烟草的转化

Construction of Plant Expression Vector of the Full Length cDNA of AlMV Replicase Gene and the Transformation of Tobacco

将苜蓿花叶病毒中国分离株(AlfalfamosaicvirusChineseisolate,A1MV-Ch)的复制酶P2亚基(90kD蛋白)基因的全长cDNA构建到植物表达载体pROK 中,得到重组植物表达载体pAlMV-FL.用三亲融合法导入农杆菌LBA4404,并转化烟草,经PCR检测,获得了含全长cDNA的转基因烟草植株.

The full length cDNA of the replicase gene which encodes 90kD proteins in the AlMV was recombinated with the plant expression vector pROKII to obtain the recombinated vector pAlMV-FL.Then the ALMV replicase gene was introduced into the tobacco cultivar via Agrobacterium tumefaciens by Triparental mating method.The PCR amplification of transgenic tobacco plants showed that the full length cDNA of replicase gene was integrated into the genome of transgenic tobacco plants.