Hahn E C, Hahn P S
Arch Virol. 1987;94(3-4):247-57. doi: 10.1007/BF01310717.
New proteins appearing after infection of cultured L929 cells with pseudorabies virus (PRV) were analyzed by SDS polyacrylamide gel electrophoresis. Analysis was facilitated by using a virus-cell system with marked inhibition of host protein synthesis after infection. Infected cells were pulsed during successive two hour periods through the infectious cycle with 35S-methionine. Proteins were extracted with detergent and analyzed on high resolution reducing gels. Thirty-four protein bands were resolved on gels of different concentrations that varied from 7 to 15 percent. Calculated apparent molecular weights of the protein peaks were not dependent on gel concentration except for very large or small sized proteins. Eight glycoproteins were resolved after labeling with 14C-glucosamine. The time course of incorporation of label was used as a measure of protein synthesis allowing the grouping of proteins according to the time of maximal synthesis. Several proteins shifted in MW during the course of infection, indicating possible post-translational cleavage or other minor modification.
用伪狂犬病病毒(PRV)感染培养的L929细胞后出现的新蛋白质,通过SDS聚丙烯酰胺凝胶电泳进行分析。使用一种病毒-细胞系统有助于分析,该系统在感染后对宿主蛋白质合成有明显抑制作用。在整个感染周期中,每隔两小时用35S-甲硫氨酸对感染细胞进行脉冲标记。用去污剂提取蛋白质,并在高分辨率还原凝胶上进行分析。在浓度从7%到15%不等的不同浓度凝胶上分辨出了34条蛋白带。除了非常大或非常小的蛋白质外,计算出的蛋白质峰的表观分子量不依赖于凝胶浓度。用14C-葡糖胺标记后分辨出了8种糖蛋白。标记掺入的时间进程被用作蛋白质合成的量度,从而可以根据最大合成时间对蛋白质进行分组。在感染过程中,几种蛋白质的分子量发生了变化,表明可能存在翻译后切割或其他微小修饰。