Triezenberg S J, Folk W R
J Virol. 1984 Aug;51(2):437-44. doi: 10.1128/JVI.51.2.437-444.1984.
We have isolated a series of point mutants in the polyomavirus origin-intergenic control region by using a procedure which exploits the single-stranded nature of DNAs cloned into M13 phage both for the generation of mutants and for their analysis by DNA sequencing. In this report we describe the effects of cytosine X guanine----thymine X adenine base-pair substitutions in the polyomavirus origin region upon replication in mouse 3T6 cells of the M13-polyomavirus constructs. Our results indicate that sequences near the center of a 34-base-pair sequence with dyad symmetry are important for replication, whereas specific nucleotides near the ends of the dyad symmetry are not important. Furthermore, a putative large T antigen-binding site at nucleotides 60 to 80 can be mutated without affecting replication function as measured in this assay.
我们通过一种利用克隆到M13噬菌体中的DNA的单链性质来产生突变体并通过DNA测序进行分析的方法，在多瘤病毒起源 - 基因间控制区域分离出了一系列点突变体。在本报告中，我们描述了多瘤病毒起源区域中胞嘧啶X鸟嘌呤----胸腺嘧啶X腺嘌呤碱基对替换对M13 - 多瘤病毒构建体在小鼠3T6细胞中复制的影响。我们的结果表明，具有二元对称的34个碱基对序列中心附近的序列对复制很重要，而二元对称末端附近的特定核苷酸则不重要。此外，在该测定中测量时，核苷酸60至80处假定的大T抗原结合位点可以发生突变而不影响复制功能。
