Rappuoli R, Ratti G, Perugini M, Murphy J R
J Virol. 1985 Apr;54(1):194-8. doi: 10.1128/JVI.54.1.194-198.1985.
A library of chromosomal DNA from Corynebacterium diphtheriae Belfanti 1030(-)tox- was cloned in the lambda phage vector EMBL4 and screened for sequences homologous to corynephage omega tox+ and the attB1-attB2 region of the C7(-)tox- chromosome. Two portions of the 1030(-)tox- chromosome, 35 and 30.5 kilobases long which contain, respectively, the entire region homologous to corynephage omega tox+ and the attB1-attB2 sites, were mapped with the restriction endonucleases BamHI and EcoRI. Chromosomal DNA from 1030(-)tox- was shown to contain a 15.5-kilobase region that was homologous to ca. 42% of the corynephage omega tox+ genome. These sequences were found to hybridize to three regions of the phage genome and do not contain either the diphtheria tox operon or the attP site. These sequences are distant from the chromosomal region that contains the attB1-attB2 sites. Moreover, unlike other known defective prophages, the physical map of this prophage starts at the cos site and is colinear with the vegetative phage map. The 30.5-kilobase region of the 1030(-)tox- chromosome, which contains the attB1-attB2 sites, has a central core region that is almost identical to the corresponding region of the C7(-)tox- chromosome; however, the flanking sequences in these two strains of C. diphtheriae are different.
将来自白喉棒状杆菌Belfanti 1030(-)tox-的染色体DNA文库克隆到λ噬菌体载体EMBL4中,并筛选与棒状噬菌体ω tox+以及C7(-)tox-染色体的attB1-attB2区域同源的序列。用限制性内切酶BamHI和EcoRI对1030(-)tox-染色体的两个部分进行了图谱绘制,这两个部分分别长35和30.5千碱基,分别包含与棒状噬菌体ω tox+同源的整个区域以及attB1-attB2位点。结果表明,来自1030(-)tox-的染色体DNA含有一个15.5千碱基的区域,该区域与棒状噬菌体ω tox+基因组的约42%同源。这些序列被发现与噬菌体基因组的三个区域杂交,并且不包含白喉毒素操纵子或attP位点。这些序列与包含attB1-attB2位点的染色体区域相距较远。此外,与其他已知的缺陷前噬菌体不同,这个前噬菌体的物理图谱从cos位点开始,并且与营养噬菌体图谱共线。1030(-)tox-染色体的30.5千碱基区域包含attB1-attB2位点,其中心核心区域与C7(-)tox-染色体的相应区域几乎相同;然而,这两株白喉棒状杆菌的侧翼序列不同。