Molecular Analysis of the TMV-Movement Protein, A Virulence Factor

植物病毒通过未知的机制在其宿主中传播 其涉及一种或多种病毒基因产物的表达。 由于病毒一般不会在其宿主中传播， 病原性，在接种植物中移动的能力 构成了毒性功能。 然而， 蛋白质和分子和细胞机制， 蛋白质的功能尚未确定。 最近这 实验室证明，烟草花叶病毒的30 kDa蛋白 病毒（TMV）负责增强细胞间的运动 利用组成型表达TMV的转基因植物， 编码30 kDa蛋白的嵌合基因。 这个优秀的系统 将用于：1）纯化和表征TMV-30 kDa 蛋白质（在此称为运动蛋白，MP）; 2）至 产生抗TMV-MP的多克隆抗体; 3） MP在转基因和TMV感染的烟草植物中的定位; 和4）对以下蛋白的功能结构域/氨基酸进行遗传作图： MP通过感染性病毒的定点和随机诱变 克隆TMV的cDNA，拯救转基因烟草上的缺陷突变体 表达MP的植物 通过分析病毒突变体 有运动缺陷，由运动能力确定 在表达30 kDa蛋白的转基因植物中， 这项研究的结果将有助于了解 TMV-30 kDa蛋白在增强细胞运动中的作用 烟草中的TMV

Plant viruses move throughout their hosts by unknown mechanisms that involve expression of one or more viral gene products. Since viruses that do not spread in their hosts are generally not pathogenic, the capacity for movement in the inoculated plant constitutes a virulence function. However, the nature of the protein(s) and the molecular and cellular mechanisms by which the proteins function are not yet determined. Recently this laboratory demonstrated that the 30 kDa protein of Tobacco Mosaic Virus (TMV) is responsible for potentiating cell-to-cell movement of TMV by using transgenic plants that constitutively express a chimeric gene encoding the 30 kDa protein. This excellent system will be used to: 1) purify and characterize the TMV-30 kDa protein (referred to here as the movement protein, MP); 2) to generate polyclonal antibodies against the TMV-MP; 3) to localize the MP in transgenic and TMV-infected tobacco plants; and 4) to genetically map the functional domains/amino acids of the MP by site directed and random mutagenesis of an infectious cloned cDNA of TMV, rescuing defective mutants on transgenic plants that express the MP. By characterizing the virus mutants that are defective in movement, as determined by ability to move in the transgenic plants that express the 30 kDa protein, the results of this research will lead to an understanding of the function of TMV-30 kDa protein in potentiating the movement of TMV in tobacco plants.