Kuo T T, Chao Y S, Lin Y H, Lin B Y, Liu L F, Feng T Y
J Virol. 1987 Jan;61(1):60-5. doi: 10.1128/JVI.61.1.60-65.1987.
It was demonstrated for the first time that filamentous bacteriophage Cflt, which contains single-stranded DNA, can incorporate its genome into that of its host. Evidence in support of the incorporation was obtained from a Southern blot hybridization analysis of DNA isolated from Cflt-lysogenized cells. DNAs from different Cflt-lysogenized cells were purified, and the integration patterns were compared. Because all integration patterns were identical and only one fragment in Cflt replicative-form DNA was missing, it appears that the integration was site specific. Only one complement of viral DNA was integrated per host chromosome. To determine the attachment site on the viral DNA, the physical map of EcoRI, XhoI, SstII, and BglII on Cflt DNA was constructed. Based on this physical map and a Southern blot hybridization analysis of lysogen DNA with these restriction endonucleases, we demonstrated that DNA sequences from all regions of the Cflt genome were represented in the integrated viral sequences. The attachment site on the viral genome was located at 69.2 to 73.8 min on the Cflt DNA.
首次证明，含有单链DNA的丝状噬菌体Cflt能够将其基因组整合到宿主基因组中。支持这种整合的证据来自对从Cflt溶源化细胞中分离出的DNA进行的Southern印迹杂交分析。纯化来自不同Cflt溶源化细胞的DNA，并比较整合模式。由于所有整合模式都是相同的，并且Cflt复制型DNA中只有一个片段缺失，因此似乎整合是位点特异性的。每个宿主染色体仅整合一份病毒DNA互补体。为了确定病毒DNA上的附着位点，构建了Cflt DNA上EcoRI、XhoI、SstII和BglII的物理图谱。基于该物理图谱以及用这些限制性内切酶对溶源DNA进行的Southern印迹杂交分析，我们证明Cflt基因组所有区域的DNA序列都存在于整合的病毒序列中。病毒基因组上的附着位点位于Cflt DNA上69.2至73.8分钟处。
