Nelson A, Roth D A, Johnson J D
Department of Molecular Biology, University of Wyoming, Laramie 82071.
Gene. 1993 May 30;127(2):227-32. doi: 10.1016/0378-1119(93)90724-h.
Antisense (AS) versions of two 51-nucleotide (nt) sequences near the 5' end of tobacco mosaic virus (TMV) RNA have been shown to inhibit in vitro translation of the adjacent gene that encodes both the 126- and 183-kDa proteins. These DNA fragments have been cloned into the binary vector, pMON530, such that either the nopaline synthase (Nos) promoter or cauliflower mosaic virus (CaMV) 35S RNA promoter is used to drive synthesis of the corresponding sense and AS RNAs. Transgenic Nicotiana tabacum cv. Xanthi nn plants containing these constructs were challenged with TMV. Plants expressing the AS orientation of a 51-nt TMV leader sequence, under the control of the CaMV 35S promoter, were found to be resistant to infection when inoculated with up to 100 times the concentration of TMV which produced severe infections in control plants. Systemic accumulation of TMV RNA and progeny virus was diminished 15 to 30-fold in these plants. Accumulation of the viral coat protein was diminished 6 to 7-fold implying a selective inhibition of TMV replication.
烟草花叶病毒（TMV）RNA 5'端附近两个51个核苷酸（nt）序列的反义（AS）版本已被证明能在体外抑制相邻基因的翻译，该基因编码126 kDa和183 kDa的蛋白质。这些DNA片段已被克隆到二元载体pMON530中，使得胭脂碱合酶（Nos）启动子或花椰菜花叶病毒（CaMV）35S RNA启动子用于驱动相应正义和反义RNA的合成。用TMV攻击含有这些构建体的转基因烟草品种Xanthi nn植株。发现在CaMV 35S启动子控制下表达51 nt TMV前导序列反义方向的植株，在接种高达对照植株严重感染浓度100倍的TMV时具有抗性。在这些植株中，TMV RNA和子代病毒的系统积累减少了15至30倍。病毒外壳蛋白的积累减少了6至7倍，这意味着对TMV复制有选择性抑制作用。
