Barnett J W, Eppstein D A, Chan H W
J Virol. 1983 Nov;48(2):384-95. doi: 10.1128/JVI.48.2.384-395.1983.
Defective herpes simplex virus type 1 genomes are composed of head-to-tail tandem repeats of small regions of the nondefective genome. Monomeric repeat units of class I defective herpes simplex virus genomes were cloned into bacterial plasmids. The repeat units functioned as replicons since both viral and convalently linked bacterial plasmid DNA replicated (with the help of DNA from nondefective virus) when transfected into rabbit skin cells. Recombinant plasmids were packaged into virions and were propagated from culture to culture by infection with progeny virus. Replication was evidently by a rolling circle mechanism since plasmid DNA was present in a high-molecular-weight form in transfected cells. Circular recombinant plasmid DNA replicated with a high degree of fidelity. In contrast, linear plasmid DNA underwent extensive deletions of both viral and bacterial sequences when transfected into rabbit skin cells. Derivative plasmids, a fraction of the size of the parental plasmid, were rescued by transforming Escherichia coli with DNA from the transfected rabbit skin cells. These plasmids functioned as shuttle vectors since they replicated faithfully in both eucaryotic and procaryotic cells.
有缺陷的单纯疱疹病毒1型基因组由无缺陷基因组小区域的头对头串联重复序列组成。I类有缺陷单纯疱疹病毒基因组的单体重复单元被克隆到细菌质粒中。这些重复单元起到复制子的作用，因为当转染到兔皮肤细胞中时，病毒和共价连接的细菌质粒DNA都会复制（在无缺陷病毒DNA的帮助下）。重组质粒被包装到病毒粒子中，并通过子代病毒感染在不同培养物中传代。复制显然是通过滚环机制进行的，因为质粒DNA在转染细胞中以高分子量形式存在。环状重组质粒DNA以高度保真度进行复制。相比之下，线性质粒DNA转染到兔皮肤细胞中时，病毒和细菌序列都会发生大量缺失。通过用转染兔皮肤细胞的DNA转化大肠杆菌，拯救出了大小为亲本质粒一部分的衍生质粒。这些质粒起到穿梭载体的作用，因为它们在真核细胞和原核细胞中都能忠实地复制。
