¹Department of Molecular Biology and Biochemistry, University of California, Irvine, CA

²Department of Genetics, University of Georgia, Athens, GA, USA

^aPresent address: Department of Molecular Microbiology and Immunology, Johns Hopkins University, Baltimore, MD, USA

^bPresent address: School of Medicine, Tufts University, Boston, MA, USA

^cPresent address: Department of Zoology, Oregon State University, Corvallis, OR, USA

Using RT-PCR, we show that mouse adenovirus type I (MAV-1) is capable of infecting and expressing in various cell types, specifically human endothelial cells. The capability of MAV-1 to infect and express in human endothelial cells makes it a potentially useful alternative to the use of human adenoviruses type 2/5 (Ad2/5) in virus-based gene therapy, although presently MAV-1 can only be produced at lower titers than Ad2/5. In this report, we present methods for the purification of MAV-1 DNA and use of this DNA along with a modified bacteria-based homologous recombination protocol to generate a full-length plasmid clone of MAV-1 DNA. Using various transfection procedures, we show that this plasmid MAV-1 DNA can generate plaques of MAV-1 virus, albeit at low efficiencies (about 0.2 p.f.u./g DNA). Furthermore, the construction of an MAV-1 plasmid along with its capability to express in human cells justifies the full development of MAV-1 into a system of gene therapy.

mouse adenovirus; human; endothelial; full-length clone