Mueller-Lantzsch N, Fan H
Cell. 1976 Dec;9(4 Pt 1):579-88. doi: 10.1016/0092-8674(76)90040-4.
A rabbit antiserum monospecific for the internal structural protein p30 of Moloney murine leukemia virus (M-MuLV) was prepared to immunoprecipitate the polyribosomes synthesizing this protein in producer cells. The antiserum was monospecific for p30 protein as judged by immunodiffusion analysis against purified p30 and total virus protein. In addition, it could specifically precipitate p30 from total virus protein in the presence of cell extracts. Less than 1% of the M-MuLV-specific messenger RNA (mRNA) could be precipitated from purified producer cell polyribosomes when the anti-p30 was used in conjunction with sheep anti-rabbit antiserum. However, considerably more virus-specific mRNA was precipitated when the anti-p30 was used in conjunction with inactivated Staphylococcus aureus, which has binding sites for the antibody. Conditions were obtained where approximately 7% of the virus-specific mRNA in purified polyribosomes was recovered by immunoprecipitation, while normal serum precipitated 10 fold less. The virus-specific mRNA in the immunoprecipitated polyribosomes was 30S-35S in size.
制备了一种针对莫洛尼鼠白血病病毒（M-MuLV）内部结构蛋白p30的兔抗血清，用于免疫沉淀产生病毒的细胞中合成该蛋白的多核糖体。通过针对纯化的p30和总病毒蛋白的免疫扩散分析判断，该抗血清对p30蛋白具有单特异性。此外，在存在细胞提取物的情况下，它可以从总病毒蛋白中特异性沉淀p30。当抗p30与羊抗兔抗血清联合使用时，从纯化的产生病毒的细胞多核糖体中沉淀出的M-MuLV特异性信使核糖核酸（mRNA）不到1%。然而，当抗p30与具有抗体结合位点的灭活金黄色葡萄球菌联合使用时，沉淀出的病毒特异性mRNA要多得多。在某些条件下，通过免疫沉淀可回收纯化多核糖体中约7%的病毒特异性mRNA，而正常血清沉淀的量则少10倍。免疫沉淀的多核糖体中的病毒特异性mRNA大小为30S-35S。
