Ebeling A, Keil G M, Knust E, Koszinowski U H
J Virol. 1983 Sep;47(3):421-33. doi: 10.1128/JVI.47.3.421-433.1983.
Murine cytomegalovirus (MCMV) Smith strain DNA is cleaved by restriction endonuclease HindIII into 16 fragments, ranging in size from 0.64 to 22.25 megadaltons. Of the 16 HindIII fragments, 15 were cloned in plasmid pACYC177 in Escherichia coli HB101 (recA). The recombinant plasmid clones were characterized by cleavage with the enzymes XbaI and EcoRI. In addition, fragments generated by double digestion of cloned fragments with HindIII and XbaI were inserted into the plasmid vector pACYC184. The results obtained after hybridization of 32P-labeled cloned fragments to Southern blots of MCMV DNA cleaved with HindIII, XbaI, EcoRI, BamHI, ApaI, ClaI, EcoRV, or KpnI allowed us to construct complete physical maps of the viral DNA for the restriction endonucleases HindIII, XbaI, and EcoRI. On the basis of the cloning and mapping experiments, it was calculated that the MCMV genome spans about 235 kilobase pairs, corresponding to a molecular weight of 155,000,000. All fragments were found to be present in equimolar concentrations, and no cross-hybridization between any of the fragments was seen. We conclude that the MCMV DNA molecule consists of a long unique sequence without large terminal or internal repeat regions. Thus, the structural organization of the MCMV genome is fundamentally different from that of the human cytomegalovirus or herpes simplex virus genome.
鼠巨细胞病毒（MCMV）史密斯株DNA被限制性内切酶HindIII切割成16个片段，大小从0.64到22.25兆道尔顿不等。在这16个HindIII片段中，有15个被克隆到大肠杆菌HB101（recA）中的质粒pACYC177中。通过用XbaI和EcoRI酶切割来鉴定重组质粒克隆。此外，将克隆片段用HindIII和XbaI双酶切产生的片段插入质粒载体pACYC184中。用32P标记的克隆片段与用HindIII、XbaI、EcoRI、BamHI、ApaI、ClaI、EcoRV或KpnI切割的MCMV DNA的Southern印迹杂交后得到的结果，使我们能够构建病毒DNA针对限制性内切酶HindIII、XbaI和EcoRI的完整物理图谱。根据克隆和图谱实验计算得出，MCMV基因组跨度约为235千碱基对，对应分子量为155,000,000。发现所有片段均以等摩尔浓度存在，且未观察到任何片段之间的交叉杂交。我们得出结论，MCMV DNA分子由一个长的独特序列组成，没有大的末端或内部重复区域。因此，MCMV基因组的结构组织与人类巨细胞病毒或单纯疱疹病毒基因组的结构组织有根本不同。
