Hightower L E, Bratt M A
J Virol. 1974 Apr;13(4):788-800. doi: 10.1128/JVI.13.4.788-800.1974.
A double-isotopic label difference analysis of polyacrylamide gels has been used to distinguish between cellular and viral protein accumulation in infected cells and to quantify the kinetics of accumulation of viral polypeptides. This technique, coupled with the determination of total radioactive amino acid incorporation in infected cultures, has revealed the following kinetic patterns. Viral polypeptides are first detected in infected cultures 2.0 to 2.5 h postinfection. The rate of accumulation of radioactive amino acids in viral polypeptides increases to a maximum (30 to 35% of the rate of accumulation in uninfected control cultures), whereas the rate of accumulation of radioactive amino acids in host-cell protein decreases to a minimum (20% of the rate of accumulation in uninfected control cultures) by 5 to 6 h postinfection. All of the viral polypeptides detected late in infection are also present at the earlier times, and the major virion structural polypeptides are present in approximately the same (N/G-2, 53K) or slightly increasing (L, G-1, M) relative amounts. One peak area containing a nonstructural glycopeptide with an apparent molecular weight of 66,000 shows significant alterations in rates of accumulation during infection. Inhibition in the rate of radioactive amino acid incorporation into both trichloroacetic acid-soluble and acid-precipitable material during infection has been demonstrated. However, these two inhibition phenomena can be uncoupled temporally by incubating infected cultures at 36 C instead of the usual 40 C, suggesting that they may not be directly related.
利用聚丙烯酰胺凝胶的双同位素标记差异分析来区分感染细胞中细胞蛋白和病毒蛋白的积累,并定量病毒多肽积累的动力学。该技术与测定感染培养物中总放射性氨基酸掺入量相结合,揭示了以下动力学模式。病毒多肽在感染后2.0至2.5小时首次在感染培养物中被检测到。病毒多肽中放射性氨基酸的积累速率增加到最大值(未感染对照培养物中积累速率的30%至35%),而宿主细胞蛋白中放射性氨基酸的积累速率在感染后5至6小时降至最小值(未感染对照培养物中积累速率的20%)。感染后期检测到的所有病毒多肽在早期也存在,主要病毒粒子结构多肽以大致相同的相对量(N/G-2、53K)或略有增加的相对量(L、G-1、M)存在。一个含有表观分子量为66,000的非结构糖肽的峰面积在感染期间积累速率有显著变化。已证明在感染期间放射性氨基酸掺入三氯乙酸可溶性和酸沉淀物质的速率均受到抑制。然而,通过将感染培养物在36℃而非通常的40℃下孵育,这两种抑制现象在时间上可以解偶联,这表明它们可能没有直接关系。