Vaquero C, Sanz A I, Serra M T, García-Luque I
Departamento de Biología de Planta, Centro de Investigaciones Biológicas (CSIC), Madrid, Spain.
Arch Virol. 1996;141(6):987-99. doi: 10.1007/BF01718603.
The complete nucleotide sequence of RNA 3 of a Spanish isolate of cucumber mosaic virus (CMV-24) has been determined. The encoded putative cell-to-cell movement protein (3a protein) and the coat protein are 279 and 218 amino acids long, respectively. The 3a protein was expressed in Escherichia coli using the vector pT7-7 and was used to raise an immunoserum. We have followed the time course of accumulation of the 3a protein, in parallel to that of the coat protein, and its subcellular localization as a function of time after CMV-24 infection on tobacco plants. The maximum accumulation level of the 3a protein was reached at early stages of infection, being detected in the cytosolic and the cell wall fractions. At later stages of infection, a decline in accumulation levels of the 3a protein was observed, and the protein was essentially associated with the cell wall fractions. These data were corroborated by immunocytochemistry performed in both infected and 3a-expressing transgenic tobacco plants.
已测定黄瓜花叶病毒西班牙分离株（CMV-24）RNA 3的完整核苷酸序列。编码的假定细胞间运动蛋白（3a蛋白）和外壳蛋白分别由279和218个氨基酸组成。使用载体pT7-7在大肠杆菌中表达3a蛋白，并用于制备免疫血清。我们追踪了3a蛋白与外壳蛋白积累的时间进程，以及CMV-24感染烟草植株后3a蛋白亚细胞定位随时间的变化。3a蛋白在感染早期达到最大积累水平，在胞质和细胞壁组分中均可检测到。在感染后期，观察到3a蛋白积累水平下降，该蛋白主要与细胞壁组分相关。在受感染和表达3a的转基因烟草植株中进行的免疫细胞化学证实了这些数据。
