Purchio A F, Erikson E, Brugge J S, Erikson R L
Proc Natl Acad Sci U S A. 1978 Mar;75(3):1567-71. doi: 10.1073/pnas.75.3.1567.
Two techniques were used to search for the polypeptide encoded by the avian sarcoma virus (ASV) src gene. First, antiserum from rabbits bearing ASV-induced fibrosarcomas was used to immunoprecipitate a transformation-specific antigen from ASV-transformed chick embryo fibroblasts. This antigen has an apparent molecular weight (Mr) of 60,000. Second, the 3' one-third of the ASV genome, selected by oligo(dT)-cellulose chromatography and sucrose gradient sedimentation, was translated in a mRNA-dependent reticulocyte cell-free lysate. This RNA species programmed the synthesis of a polypeptide that comigrated with the transformation-specific antigen of Mr 60,000 immunoprecipitated from transformed cells. The methionine-containing tryptic peptides from the polypeptides of Mr 60,000 obtained from translation in vitro and from immunoprecipitation were found to be identical upon two-dimensional fractionation.
采用了两种技术来寻找禽肉瘤病毒（ASV）src基因编码的多肽。首先，用携带ASV诱导的纤维肉瘤的兔子的抗血清从ASV转化的鸡胚成纤维细胞中免疫沉淀一种转化特异性抗原。这种抗原的表观分子量（Mr）为60,000。其次，通过寡聚（dT）-纤维素色谱法和蔗糖梯度沉降法选择的ASV基因组的3'三分之一在依赖mRNA的网织红细胞无细胞裂解物中进行翻译。这种RNA种类指导合成一种与从转化细胞中免疫沉淀的60,000 Mr的转化特异性抗原迁移率相同的多肽。在二维分级分离后发现，从体外翻译和免疫沉淀获得的60,000 Mr多肽的含甲硫氨酸的胰蛋白酶肽是相同的。
