Ito Y, Spurr N, Dulbecco R
Proc Natl Acad Sci U S A. 1977 Mar;74(3):1259-63. doi: 10.1073/pnas.74.3.1259.
High-titer antiserum raised in rats against the tumor (T) antigen of polyoma virus was used to purify the T antigen by the Staphylococcus protein A antibody adsorbent technique. Sodium dodecyl sulfate/polyacrylamide gel electrophoresis allowed the identification of a protein with an apparent molecular weight of 100,000-108,000 as a major component induced in lytically infected mouse cells. In cells infected by ts A mutants this component was temperature sensitive. Several minor components were also observed. In pulse and chase experiments there was a slight decrease in electrophoretic mobility of T antigen during the chase period at the permissive temperature, suggesting that the T antigen is a modified protein. In two lines of transformed cells, the amount of T antigen seemed to be considerably less than in lytically infected cells, but the size of the antigen appeared to be equal.
用在大鼠体内产生的针对多瘤病毒肿瘤(T)抗原的高效价抗血清,通过葡萄球菌蛋白A抗体吸附技术来纯化T抗原。十二烷基硫酸钠/聚丙烯酰胺凝胶电泳能够鉴定出一种表观分子量为100,000 - 108,000的蛋白质,它是在裂解感染的小鼠细胞中诱导产生的主要成分。在被ts A突变体感染的细胞中,该成分对温度敏感。还观察到了几种次要成分。在脉冲追踪实验中,在允许温度下的追踪期内,T抗原的电泳迁移率略有下降,这表明T抗原是一种经过修饰的蛋白质。在两株转化细胞系中,T抗原的量似乎比裂解感染的细胞中少得多,但抗原的大小似乎相等。