Cordier O, Tondre L, Aubertin A M, Kirn A
Virology. 1986 Jul 30;152(2):355-64. doi: 10.1016/0042-6822(86)90138-8.
Multiplication of frog virus 3 (FV 3) occurs in mammalian cells provided they are incubated at temperatures lower than 33 degrees. The expression of the viral genome at supraoptimal temperatures was followed by analyzing the polypeptides produced in CHO-infected cells and comparing with those obtained under restrictive conditions provoked by amino acid analogs or metabolic inhibitors. Late polypeptides were not detected at 33 degrees and the number of delayed early species decreased gradually with increasing temperatures consequently the synthesis of all delayed early proteins was not turned on in response to a unique event. At 37 degrees the synthesis was limited to the immediate early species, i.e., the proteins synthesized after cycloheximide reversal. Temperature shift experiments suggested that delayed early genes remained untranscribed at 37 degrees. Thus, incubation of FV 3-infected mammalian cells at 37 degrees provides a unique way of limiting viral synthesis to immediate early proteins without the side-effect provoked by inhibitors.
蛙病毒3(FV 3)可在哺乳动物细胞中增殖,前提是将这些细胞在低于33摄氏度的温度下培养。通过分析受CHO感染的细胞中产生的多肽,并与在氨基酸类似物或代谢抑制剂引发的限制条件下获得的多肽进行比较,对病毒基因组在超适宜温度下的表达进行了追踪。在33摄氏度时未检测到晚期多肽,随着温度升高,延迟早期蛋白的数量逐渐减少,因此,并非所有延迟早期蛋白的合成都是由一个独特事件开启的。在37摄氏度时,合成仅限于即刻早期蛋白,即放线菌酮作用逆转后合成的蛋白。温度转换实验表明,延迟早期基因在37摄氏度时仍未转录。因此,将感染FV 3的哺乳动物细胞在37摄氏度下培养,提供了一种将病毒合成限制为即刻早期蛋白的独特方法,且不会产生抑制剂引发的副作用。