Fleissner E, Tress E
J Virol. 1973 Feb;11(2):250-62. doi: 10.1128/JVI.11.2.250-262.1973.
Several methods have been explored for the detection and characterization of viral proteins from soluble extracts of cells transformed by Rous sarcoma virus (RSV). Viral antigens have been analyzed after gel filtration in several solvents. In addition, immune complexes formed with virus-specific sera have been isolated by agarose gel filtration and by high- or low-speed centrifugation through sucrose solutions. Radioactive proteins from these immune complexes have been analyzed by gel filtration in 6 m guanidine hydrochloride or by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. Comparison with proteins from purified virus indicates the presence of two viral core proteins (gs1 and gs2) in the soluble fraction from virus-producing chicken cells. In the same fraction from RSV-transformed hamster cells (which do not produce virus), three gs proteins (gs1, gs2, and gs3) could be identified. The soluble viral gs proteins are strongly bound to at least two larger polypeptides in cell extracts. These polypeptides do not appear to be viral in origin and have the property of undergoing a time-dependent aggregation in the extracts. One of these cell-derived proteins, which is present in a variety of uninfected cell types, closely resembles actin.
人们已经探索了多种方法来检测和鉴定劳斯肉瘤病毒(RSV)转化细胞的可溶性提取物中的病毒蛋白。病毒抗原在几种溶剂中进行凝胶过滤后进行了分析。此外,与病毒特异性血清形成的免疫复合物已通过琼脂糖凝胶过滤以及通过蔗糖溶液进行高速或低速离心分离出来。这些免疫复合物中的放射性蛋白已通过在6M盐酸胍中进行凝胶过滤或在十二烷基硫酸钠存在下进行聚丙烯酰胺凝胶电泳进行分析。与来自纯化病毒的蛋白质进行比较表明,在产生病毒的鸡细胞的可溶性部分中存在两种病毒核心蛋白(gs1和gs2)。在RSV转化的仓鼠细胞(不产生病毒)的同一部分中,可以鉴定出三种gs蛋白(gs1、gs2和gs3)。可溶性病毒gs蛋白在细胞提取物中与至少两种较大的多肽紧密结合。这些多肽似乎不是病毒来源的,并且具有在提取物中随时间发生聚集的特性。这些细胞衍生蛋白之一存在于多种未感染的细胞类型中,与肌动蛋白非常相似。