Jørgensen P, Mikkelsen T, Pedersen K, Pedersen F S, Kjeldgaard N O
Institute of Molecular Biology, Aarhus University, Denmark.
Gene. 1991 Dec 30;109(2):243-7. doi: 10.1016/0378-1119(91)90615-i.
The bacterial lactose operator (lacO) was introduced into the PstI site of the long terminal repeat of the SL3-3 murine leukemia virus, generating a virus, SL3-3lacO, that can replicate in NIH3T3 cell cultures. DNA sequences harboring the lacO sequence might be recovered by molecular cloning in Escherichia coli lac+ lacZ+ using bacteriophage lambda or plasmid vectors. The high copy numbers of the lacO sequence titrate out the lac repressor, leading to the induction of the lac operon in the host. We show here that the lacO and the proviral sequences are carried stably together in the genomes of SL3-3lacO-infected cell cultures and in viral particles. This system is designed to facilitate studies on the provirus and the site of viral integration.
将细菌乳糖操纵基因(lacO)导入SL3-3鼠白血病病毒长末端重复序列的PstI位点,产生了一种能在NIH3T3细胞培养物中复制的病毒,即SL3-3lacO。携带lacO序列的DNA片段可通过使用噬菌体λ或质粒载体在大肠杆菌lac+ lacZ+中进行分子克隆来回收。lacO序列的高拷贝数会使lac阻遏物失活,从而导致宿主中lac操纵子的诱导。我们在此表明,lacO和前病毒序列在SL3-3lacO感染的细胞培养物基因组和病毒颗粒中稳定共存。该系统旨在促进对前病毒和病毒整合位点的研究。