a selectable and excisable marker system for the rapid creation of recombinant poxviruses可选择的,可割取的重组痘病毒的快速创建标记系统.pdfVIP

A Selectable and Excisable Marker System for the Rapid Creation of Recombinant Poxviruses 1,2. 2. 3 3 2 Julia L. Rintoul , Jiahu Wang , Don B. Gammon , Nicholas J. van Buuren , Kenneth Garson , Karen 2 3 3 1,2 Jardine , Michele Barry , David H. Evans , John C. Bell * 1 Department of Biochemistry, Microbiology and Immunology, Faculty of Medicine, University of Ottawa, Ottawa, Canada, 2 Centre for Cancer Therapeutics, Ottawa Hospital Research Institute, Ottawa, Canada, 3 Department of Medical Microbiology and Immunology, Faculty of Medicine and Dentistry, Li Ka Shing Institute of Virology, University of Alberta, Edmonton, Canada Abstract Background: Genetic manipulation of poxvirus genomes through attenuation, or insertion of therapeutic genes has led to a number of vector candidates for the treatment of a variety of human diseases. The development of recombinant poxviruses often involves the genomic insertion of a selectable marker for purification and selection purposes. The use of marker genes however inevitably results in a vector that contains unwanted genetic information of no therapeutic value. Methodology/Principal Findings: Here we describe an improved strategy that allows for the creation of marker-free recombinant poxviruses of any species. The Selectable and Excisable Marker (SEM) system incorporates a unique fusion marker gene for the efficient selection of poxvirus recombinants and the Cre/loxP system to facilitate the subsequent removal of the marker. We have defined and characterized this new methodological tool by insertion of a foreign gene