van Dun C M, Bol J F, Van Vloten-Doting L
Department of Biochemistry, State University of Leiden, P. O. Box 9505, 2300 RA Leiden, The Netherlands.
Virology. 1987 Aug;159(2):299-305. doi: 10.1016/0042-6822(87)90467-3.
Using the Agrobacterium tumefaciens binary vector system, a chimeric gene consisting of the cauliflower mosaic virus 35 S promoter, alfalfa mosaic virus (AIMV) coat protein (CP) cistron, and the nopaline synthase polyadenylation signal was integrated into the genome of Nicotiana tabacum cv. Samsun NN. In 70% of the transgenic tobacco plants the chimeric mRNA and its translation product could be detected. CP accumulated to levels up to 0.05% of the soluble leaf protein. The accumulation was independent of leaf age. The same approach was undertaken for the CP of tobacco rattle virus (TRV). The chimeric gene was integrated in the genome of Nicotiana tabacum cv. Xanthi nc. The results with respect to the accumulation of the chimeric mRNA and TRV CP in leaves of transgenic tobacco plants were comparable to those with AIMV transformed plants. Plants accumulating AIMV CP were highly resistant to infection with AIMV nucleoproteins but could be infected with a mixture of AIMV RNAs 1-4. Moreover, a mixture of AIMV RNAs 1, 2, and 3 was infectious to these plants but not to nontransformed control plants.
利用根癌农杆菌双元载体系统，将一个由花椰菜花叶病毒35S启动子、苜蓿花叶病毒（AIMV）外壳蛋白（CP）顺反子和胭脂碱合酶聚腺苷酸化信号组成的嵌合基因整合到烟草品种Samsun NN的基因组中。在70%的转基因烟草植株中可检测到嵌合mRNA及其翻译产物。CP积累量高达可溶性叶蛋白的0.05%。这种积累与叶龄无关。对烟草脆裂病毒（TRV）的CP也采用了同样的方法。嵌合基因整合到烟草品种Xanthi nc的基因组中。转基因烟草植株叶片中嵌合mRNA和TRV CP积累的结果与AIMV转化植株的结果相当。积累AIMV CP的植株对AIMV核蛋白感染具有高度抗性，但可被AIMV RNA 1-4的混合物感染。此外，AIMV RNA 1、2和3的混合物对这些植株具有感染性，但对未转化的对照植株没有感染性。
