Tan B T, Wu L, Berk A J
Department of Microbiology and Molecular Genetics, Molecular Biology Institute, University of California, Los Angeles, Los Angeles, California 90095, USA.
J Virol. 1999 Sep;73(9):7582-9. doi: 10.1128/JVI.73.9.7582-7589.1999.
EBV episomes are nuclear plasmids that are stably maintained through multiple cell divisions in primate and canine cells (J. L. Yates, N. Warren, and B. Sugden, Nature 313:812-815, 1985). In this report, we describe the construction and characterization of an E1-deleted recombinant adenovirus vector system that delivers an EBV episome to infected cells. This adenovirus-EBV hybrid vector system utilizes Cre-mediated, site-specific recombination to excise an EBV episome from a target recombinant adenovirus genome. We demonstrate that this vector system efficiently delivers the EBV episome and stably transforms a large fraction of infected canine D-17 cells. Using a colony-forming assay, we demonstrate stable transformation of 37% of cells that survive the infection. However, maximal transformation efficiency is achieved at doses of the E1-deleted recombinant adenoviruses that are toxic to the infected cells. Consequently, E1-deleted vector toxicity imposes a limitation on our current vector system.
EBV附加体是核质粒，可在灵长类和犬类细胞的多次细胞分裂中稳定维持（J.L.耶茨、N.沃伦和B.萨格登，《自然》313:812 - 815，1985）。在本报告中，我们描述了一种E1缺失的重组腺病毒载体系统的构建和特性，该系统将EBV附加体递送至感染细胞。这种腺病毒 - EBV杂交载体系统利用Cre介导的位点特异性重组从目标重组腺病毒基因组中切除EBV附加体。我们证明该载体系统能有效地递送EBV附加体，并稳定转化大部分感染的犬D - 17细胞。通过集落形成试验，我们证明在感染后存活的细胞中有37%实现了稳定转化。然而，在对感染细胞有毒性的E1缺失重组腺病毒剂量下可实现最大转化效率。因此，E1缺失载体的毒性对我们当前的载体系统构成了限制。
