van Dun C M, van Vloten-Doting L, Bol J F
Department of Biochemistry, State University of Leiden, The Netherlands.
Virology. 1988 Apr;163(2):572-8. doi: 10.1016/0042-6822(88)90298-x.
Chimeric genes composed of DNA complementary to alfalfa mosaic virus (AIMV) RNAs 1 or 2, the CaMV 35 S promoter, and the nos polyadenylation signal were transferred to the genome of Nicotiana tabacum cv. Samsun NN by means of the Agrobacterium tumefaciens transformation system. Transformants contained intact copies of the viral genes and accumulated transcripts of approximately the size predicted from the cloning procedure. Using antisera raised against synthetic peptides corresponding to the C-terminal parts of AIMV P1 and P2, it was not possible to detect viral translation products in the transformants. However, transgenic protoplasts containing cDNA1 were able to complement an infection by the AIMV nucleoproteins containing RNAs 2 and 3, indicating that biologically active P1 accumulates in these protoplasts. Upon inoculation with AIMV strains 425 or YSMV, the cDNA1- and cDNA2-transformed plants became infected to a level similar to that of nontransformed or vector-transformed control plants.
由与苜蓿花叶病毒(AIMV)RNA 1或2互补的DNA、花椰菜花叶病毒35S启动子和胭脂碱合酶基因(nos)多聚腺苷酸化信号组成的嵌合基因,通过根癌农杆菌转化系统转移到烟草品种Samsun NN的基因组中。转化体含有病毒基因的完整拷贝,并积累了大小与克隆过程预测相近的转录本。使用针对与AIMV P1和P2 C端部分对应的合成肽产生的抗血清,无法在转化体中检测到病毒翻译产物。然而,含有cDNA1的转基因原生质体能够互补由含有RNA 2和3的AIMV核蛋白引发的感染,这表明具有生物活性的P1在这些原生质体中积累。用AIMV菌株425或YSMV接种后,cDNA1和cDNA2转化的植物被感染的程度与未转化或载体转化的对照植物相似。