caa a22 4500 445823275 CHVBK 20180317145251.0 cr unu---uuuuu 170323e20110801xx s 000 0 eng 10.1007/s10616-011-9357-6 doi (NATIONALLICENCE)springer-10.1007/s10616-011-9357-6 A single plasmid transfection that offers a significant advantage associated with puromycin selection, fluorescence-assisted cell sorting, and doxycycline-inducible protein expression in mammalian cells [Elektronische Daten] [Takayuki Iwaki, Kazuo Umemura] Although there are several inducible expression systems for mammalian cells, the most reliable one is the tetracycline-regulated expression system. This system is well-established and widely used by many researchers. Although Clontech provides several types of cells that stably express reverse tetracycline transactivator (rtTA), the cells that are not provided can be generated with pTet-On-Advanced by first integrating this plasmid into the require type of cell and then introducing the genes of interest. These processes are experimental bottlenecks. To improve this situation, we synthesized an all-in-one vector, termed pMAK17, which enables constitutive expression of puromycin N-acetyltransferase, modified Discosoma red fluorescent protein, and rtTA, as well as PTight-driven enhanced green fluorescent protein (EGFP). The pMAK17-transfected cells could be successfully induced to express EGFP, were selectable by fluorescence-activated cell sorting, and displayed puromycin resistance. Springer Science+Business Media B.V., 2011 DsRED2 nationallicence Inducible nationallicence rtTA nationallicence Puromycin nationallicence Selection nationallicence Single plasmid nationallicence Iwaki Takayuki Department of Pharmacology, Hamamatsu University School of Medicine, 1-20-1 Handayama, Hamamatsu, 431-3192, Shizuoka, Japan aut Umemura Kazuo Department of Pharmacology, Hamamatsu University School of Medicine, 1-20-1 Handayama, Hamamatsu, 431-3192, Shizuoka, Japan aut Cytotechnology Springer Netherlands 63/4(2011-08-01), 337-343 0920-9069 63:4<337 2011 63 10616 https://doi.org/10.1007/s10616-011-9357-6 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10616-011-9357-6 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Iwaki Takayuki Department of Pharmacology, Hamamatsu University School of Medicine, 1-20-1 Handayama, Hamamatsu, 431-3192, Shizuoka, Japan aut NATIONALLICENCE 700 1- Umemura Kazuo Department of Pharmacology, Hamamatsu University School of Medicine, 1-20-1 Handayama, Hamamatsu, 431-3192, Shizuoka, Japan aut NATIONALLICENCE 773 0- Cytotechnology Springer Netherlands 63/4(2011-08-01), 337-343 0920-9069 63:4<337 2011 63 10616 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer