禽成髓细胞瘤病毒和劳氏鼠白血病病毒每个病毒体的逆转录酶活性。
Reverse transcriptase activity per virion for avian myeloblastosis virus and Rauscher murine leukemia virus.
作者信息
Liebes L F, Rich M A, McCormick J J, Salmeen I, Rimai L
出版信息
J Virol. 1976 Apr;18(1):42-7. doi: 10.1128/JVI.18.1.42-47.1976.
We have measured reverse transcriptase enzyme activity per virus particle for samples of avian myeloblastosis virus (BAI strain) and murine leukemia virus (RAUSCHER) USing the synthetic template poly(rC)-oligo(dG). Absolute virus concentrations were determined directly by laser beat frequency spectroscopy. Enzyme activity per virion was determined from the slope of the activity plotted as a function of virus concentration. With this reverse transcriptase assay, the minimum activity (expressed as picomoles of dGTP incorporated/virion per hour) is estimated at (28.1 +/- 4.2) X 10(-7) for avian myeloblastosis virus and (1.1 +/- 0.2) X 10(-7) for murine leukemia virus. The sensitivity of this assay, which is determined by the level of incorporated radioactivity measurable above background, is 2.5 X 10(-4) virions for avian myeloblastosis virus (with dGTP specific activity of 8.9 Ci/mmol) and 88 X 10(-4) virions for murine leukemia virus (with dGTP specific activity of 6.52 CI/mmol). These results show that although reverse transcriptase assays can obviously be used to measure relative virus concentrations of equally purified samples of the same virus, they can be very misleading when used to compare the concentrations of different virus species.
我们使用合成模板聚（rC）-寡聚（dG）测量了禽成髓细胞瘤病毒（BAI株）和鼠白血病病毒（RAUSCHER）样本中每个病毒颗粒的逆转录酶活性。通过激光拍频光谱法直接测定绝对病毒浓度。每个病毒体的酶活性由活性与病毒浓度的函数关系图的斜率确定。通过这种逆转录酶测定法，禽成髓细胞瘤病毒的最低活性（以每小时每个病毒体掺入的dGTP皮摩尔数表示）估计为（28.1±4.2）×10⁻⁷，鼠白血病病毒为（1.1±0.2）×10⁻⁷。该测定法的灵敏度由高于背景可测量的掺入放射性水平决定，对于禽成髓细胞瘤病毒（dGTP比活为8.9 Ci/mmol）为2.5×10⁻⁴个病毒体，对于鼠白血病病毒（dGTP比活为6.52 Ci/mmol）为88×10⁻⁴个病毒体。这些结果表明，虽然逆转录酶测定法显然可用于测量同一病毒的同等纯化样本的相对病毒浓度，但用于比较不同病毒种类的浓度时可能会产生很大误导。
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