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Systemic expression of a bacterial gene by a tobacco mosaic virus-based vector.

机译:基于烟草花叶病毒的载体对细菌基因的系统表达。

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摘要

Tobacco mosaic virus (TMV) produces large quantities of RNA and protein on infection of plant cells. This and other features, attributable to its autonomous replication, make TMV an attractive candidate for expression of foreign sequences in plants. However, previous attempts to construct expression vectors based on plant RNA viruses, such as TMV, have been unsuccessful in obtaining systemic and stable movement of foreign genes to uninoculated leaves in whole plants. A hybrid viral RNA (TB2) was constructed, containing sequences from two tobamoviruses (TMV-U1 and odontoglossum ringspot virus). Two bacterial sequences inserted independently into TB2 moved systemically in Nicotiana benthamiana, although they differed in their stability on serial passage. Systemic expression of the bacterial protein neomycin phosphotransferase was demonstrated. Hybrid RNAs containing both TMV-U1 and the inserted bacterial gene sequences were encapsidated by the odontoglossum ringspot virus coat protein, facilitating their transmission and amplification on passaging to subsequent plants. The vector TB2 provides a rapid means of expressing genes and gene variants in plants.
机译:烟草花叶病毒(TMV)在感染植物细胞后会产生大量RNA和蛋白质。归因于它的自主复制,这种和其他特征使TMV成为在植物中表达外源序列的有吸引力的候选者。然而,先前基于植物RNA病毒如TMV构建表达载体的尝试未能获得整株植物中外源基因向未接种叶片的系统和稳定的运动。构建了一个杂种病毒RNA(TB2),其中包含来自两种Tobamoviruses(TMV-U1和齿don圆环斑病毒)的序列。独立插入到TB2中的两个细菌序列在本氏烟草中系统地移动,尽管它们在连续传代时的稳定性不同。证明了细菌蛋白新霉素磷酸转移酶的系统表达。牙本质环斑病毒外壳蛋白将同时含有TMV-U1和插入的细菌基因序列的杂合RNA衣壳化,从而促进它们在传代到随后的植物中的传播和扩增。载体TB2提供了在植物中表达基因和基因变体的快速手段。

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