Mikke R, Kielanowska M, Shugar D, Zmudzka B
Nucleic Acids Res. 1976 Jun;3(6):1603-11. doi: 10.1093/nar/3.6.1603.
Poly(2'-O-ethylcytidylate) is a poor template-primer for purified avian myeloblastosis virus reverse transcriptase; the relative activities of the template-primers poly(C)-oligo(dG), poly(Cm)-oligo(dG) and poly(Ce)-oligo(dG) are 23:16:1. A mixture of poly(Ce) and poly(dI) is inactive as template-primer, in agreement with the observed inability of these to form a helical complex. By contrast the inactivity of poly(Ce)-poly(I) is shown to be due to the influence of the 2'-O-ethyl residue. Poly(Ce) inhibits poly(A)-oligo(dT)-directed polymerase activity, with Ki = 3 muM, but marked inhibition with poly(A)-poly(dT) occurs only at low concentrations of the latter. Poly(Ce) did not inhibit template-primer activity of poly(C)-poly(dI) and poly(dC)-poly(dI). Qualitative physico-chemical studies show only partial complex formation between oligo(dG) and poly(C) and its 2'-O-alkyl analogues. This is discussed in relation to the widespread use of poly(C)-oligo(dG) as the template-primer for reverse transcriptase.
聚(2'-O-乙基胞苷酸)对于纯化的禽成髓细胞瘤病毒逆转录酶而言是一种较差的模板引物;模板引物聚(C)-寡聚(dG)、聚(Cm)-寡聚(dG)和聚(Ce)-寡聚(dG)的相对活性为23:16:1。聚(Ce)和聚(dI)的混合物作为模板引物无活性,这与观察到的它们无法形成螺旋复合物一致。相比之下,聚(Ce)-聚(I)的无活性表明是由于2'-O-乙基残基的影响。聚(Ce)抑制聚(A)-寡聚(dT)指导的聚合酶活性,Ki = 3 μM,但仅在聚(A)-聚(dT)浓度较低时才会出现明显抑制。聚(Ce)不抑制聚(C)-聚(dI)和聚(dC)-聚(dI)的模板引物活性。定性物理化学研究表明,寡聚(dG)与聚(C)及其2'-O-烷基类似物之间仅形成部分复合物。这与聚(C)-寡聚(dG)作为逆转录酶模板引物的广泛使用相关进行了讨论。