Panet A, Baltimore D, Hanafusa T
J Virol. 1975 Jul;16(1):146-52. doi: 10.1128/JVI.16.1.146-152.1975.
A radioimmunoassay was developed that can detect and quantitate 3 ng or more of the avian RNA tumor virus reverse transcriptase. The assay detected no antigenic sites in Rous sarcoma virus alpha virions or in virions of a murine RNA tumor virus. About 70 molecules of reverse transcriptase were found per virion of avian myleloblastosis virus with this assay or with an assay based on antibody inhibition of enzymatic activity. The assay detected about 270 ng of enzyme per mg of cell protein in virus-producing cells; uninfected cells had much less antigenic material but contained some determinants able to displace radioactive antigen. No additional antigenic determinants on reverse transcriptase could be detected that were not found on the separated alpha subunit of the enzyme. Although sevenfold less sensitive than enzymatic activity as a measure of reverse transcriptase, the radioimmunoassay can detect antigen using small amounts of protein and in the presence of inhibtors.
已开发出一种放射免疫测定法,该方法能够检测并定量3纳克或更多的禽RNA肿瘤病毒逆转录酶。该测定法在劳氏肉瘤病毒α病毒粒子或鼠RNA肿瘤病毒的病毒粒子中未检测到抗原位点。用该测定法或基于抗体抑制酶活性的测定法,发现每一个禽成髓细胞瘤病毒粒子中约有70个逆转录酶分子。该测定法在产生病毒的细胞中每毫克细胞蛋白检测到约270纳克的酶;未感染的细胞抗原物质少得多,但含有一些能够置换放射性抗原的决定簇。在该酶分离出的α亚基上未发现的逆转录酶上的额外抗原决定簇无法被检测到。尽管作为一种测量逆转录酶的方法,放射免疫测定法的灵敏度比酶活性低七倍,但它能够在存在抑制剂的情况下使用少量蛋白质检测抗原。