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pQuattro vectors allow one-step multigene metabolic engineering and auto-selection of quattrocistronic artificial mammalian operons

Fussenegger, Martin ; Moser, Samuel ; Bailey, James

In: Cytotechnology, 1998, vol. 28, no. 1-3, p. 229-235

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    Titre
    • pQuattro vectors allow one-step multigene metabolic engineering and auto-selection of quattrocistronic artificial mammalian operons
    Auteur
    • Fussenegger, Martin. Swiss Federal Institute of Technology, ETH Zurich, Institute of Biotechnology, CH-8093, Zurich, Switzerland
    • Moser, Samuel. Swiss Federal Institute of Technology, ETH Zurich, Institute of Biotechnology, CH-8093, Zurich, Switzerland
    • Bailey, James. Swiss Federal Institute of Technology, ETH Zurich, Institute of Biotechnology, CH-8093, Zurich, Switzerland
    Type de document
    • Postprint
    Langue
    • Anglais
    Publié dans
    • Cytotechnology, 1998, vol. 28, no. 1-3, p. 229-235. Kluwer Academic Publishers
    Autre version électronique
    Identifiant OAI-PMH
    • oai:doc.rero.ch:322162
    Summary
    Based on internal ribosomal entry sites (IRES) of picornaviral origin we constructed a novel family of mammalian expression vectors. pQuattro vectors contain quattrocistronic artificial eukaryotic operons which link, in a single transcript, the simultaneous and coordinated as well as adjustable expression of up to three independent genes of interest to a terminal neomycin (neo) resistance marker. Due to the strict genetic linkage of the transgenes and the terminal selection marker, this genetic configuration enables, by the selection on neomycin, multigene metabolic engineering of mammalian cells in a single step (one-step metabolic engineering). Furthermore, selection on the terminal cistron of multicistronic expression units enforces cocistronic expression of all upstream encoded genes and maximises genetic integrity of the eukaryotic operon in stable mammalian cell lines, since clones harbouring damaged multicistronic expression units become neomycin-sensitive and are automatically counterselected (auto-selection). The modular set-up and the abundance of restriction sites in pQuattro vectors facilitate the movement of individual genes between multicistronic expression vectors and guarantees high compatibility with genetic elements of a wide variety of existing mammalian expression vectors