Wu A M, Cetta A
Biochemistry. 1975 Feb 25;14(4):789-95. doi: 10.1021/bi00675a022.
Nonionic detergents stimulate purified RNA-directed DNA polymerase (reverse transcriptase) activity from various RNA tumor viruses ranging from avian to primate species. The stimulatory effect of the nonionic detergent is dependent on the type and amount of template-primer. The greatest stimulation is obtained when high concentrations of (dT)12-18-(rA)n or activated salmon sperm DNA are used as template-primers. Little stimulation is obtained with viral 70S RNA or with (dT)12-18- (dA)n. The detergent stimulation appears to be specific for viral reverse transcriptase since this effect is not observed with purified bacterial DNA polymerase or with three known mammalian cellular DNA polymerases. This finding may, therefore, be a useful additional criterion for distinguishing viral reverse transcriptase isolated from cells from other cellular DNA polymerases. Nonionic detergent also has a stabilizing effect on viral DNA polymerase against thermal inactivation. This stabilizing effect is further enhanced by the presence of template-primer.
非离子去污剂可刺激从禽类到灵长类等多种RNA肿瘤病毒的纯化RNA指导的DNA聚合酶(逆转录酶)活性。非离子去污剂的刺激作用取决于模板引物的类型和数量。当使用高浓度的(dT)12-18-(rA)n或活化的鲑鱼精DNA作为模板引物时,可获得最大的刺激效果。使用病毒70S RNA或(dT)12-18-(dA)n时,刺激作用很小。去污剂的刺激作用似乎对病毒逆转录酶具有特异性,因为在纯化的细菌DNA聚合酶或三种已知的哺乳动物细胞DNA聚合酶中未观察到这种效应。因此,这一发现可能是区分从细胞中分离出的病毒逆转录酶与其他细胞DNA聚合酶的一个有用的额外标准。非离子去污剂对病毒DNA聚合酶还具有抗热失活的稳定作用。模板引物的存在可进一步增强这种稳定作用。