title: An autocatalytic expression system for regulated production of recombinant protein in mammalian cells.
abstract: An autocatalytic inducible mammalian expression system was established. This system is composed of two sets of vectors: one carries a selectable marker gene and the other carries a bicistronic expression unit consisting of a target gene, an internal ribosomal entry site, and a tetracycline-controlled transactivator gene. Both the selectable marker and the bicistronic unit are controlled by the tetracycline-responsive promoter (PhCMV*-1). When the two vectors are cotransfected into host cells, only a single selection round in the absence of tetracycline is necessary to generate clones expressing the target gene. The expression level is high and easily regulated by tetracycline. Combination with a gene amplification system may allow further enhance the foreign gene expression. Because the PhCMV*-1 promoter is relatively independent of cellular regulation signals, this system is expected to work in a wide variety of cell types.
authors.affiliation: Graduate School of Genetic Resources Technology, Kyushu University, Fukuoka, 812-81, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Zhang
authors.forename: Y
authors.initials: Y
authors.lastname: Katakura
authors.forename: H
authors.initials: H
authors.lastname: Ohashi
authors.forename: S
authors.initials: S
authors.lastname: Shirahata
mesh_terms: D000818:Animals; D016466:CHO Cells; D002384:Catalysis; D006224:Cricetinae; D004797:Enzyme-Linked Immunosorbent Assay; D005784:Gene Amplification; D015870:Gene Expression; D005796:Genes; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors; D006801:Humans; D015850:Interleukin-6 / Q000096:biosynthesis / Q000235:genetics; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000096:biosynthesis*; D015183:Restriction Mapping; D013752:Tetracycline / Q000494:pharmacology; D015533:Transcriptional Activation / Q000187:drug effects; D014162:Transfection
chemical_list: D015850:Interleukin-6; D011994:Recombinant Proteins; D013752:Tetracycline