【摘要】 根据烟草花叶病毒蚕豆株系 (TMV- B)的已知序列合成引物 ,经 PCR扩增获得 TMV- B的移动蛋白 (MP)基因 .该基因测序验证后 ,分别以正、反向克隆到植物表达载体 p BI12 1中 ,并置于 Ca MV35S启动子控制之下 ,通过三亲交配及叶盘转化 ,将 MP基因导入烟草 .经卡那霉素抗性筛选 ,PCR扩增 ,Southern点杂交 ,Northern点杂交及 Western blot检测 ,获得了正义表达 MP基因及反义表达 MP基因的转基因烟草 ,分别命名为 p TMP(+ )烟草和 p TMP(- )烟草更多还原

【Abstract】 ]The movement protein (MP) gene of tobacco mosaic virus isolated from faba bean was amplified by RT PCR technique with specific primers designed on the basis of reported TMV B sequence. The cDNA fragment was cloned into pGEM T easy vector and then sequenced. The sequence was identical with reported TMV B MP sequence. The MP gene was inserted into plant expression vector pBI121 in viral sense [pTMP(+)] and antisense [pTMP(-)], under the control of CaMV 35S promoter. Both of pTMP(+) and pTMP(-) DNA was transformed into Nicotiana tabacum by Agrobacterium tumefaciens LBA4404. PCR, Southern dot blot, Northern dot blot and Western blot confirmed MP genes were expressed in viral sense and antisense in transgenic plants.更多还原