An autocatalytic expression system for regulated production of recombinant protein in mammalian cells.
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.
Zhang
Y
Y
Graduate School of Genetic Resources Technology, Kyushu University, Fukuoka, 812-81, Japan.
Katakura
Y
Y
Ohashi
H
H
Shirahata
S
S
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
D015850:Interleukin-6; D011994:Recombinant Proteins; D013752:Tetracycline