title: Inducible protein expression in Drosophila Schneider 2 cells using the lac operator-repressor system.
abstract: Schneider line 2 cells, derived from Drosophila melanogaster, can be used as a highly versatile gene expression system. Two powerful promoters derived from the actin5C (Ac5) and metallothionein (Mtn) genes are available. The Mtn promoter can be used for the inducible expression of heterologous proteins unsuitable for constitutive expression. However, to circumvent using CuSO(4) or CdCl(2) as inducers of the Mtn promoter, we created a modified Ac5 promoter, Ac5LacO, in which two short lac operator sequences are embedded. Expression from the Ac5LacO promoter was regulated with co-expression of the lac repressor and IPTG. More than 25-fold induction of firefly luciferase expression was achieved in transient transfection experiments. Furthermore, we demonstrated that the lac operator-repressor regulatory system functioned in chromosomally integrated cell lines.
authors.affiliation: RIKEN Systems and Structural Biology Center, Tsurumi, Yokohama 230-0045, Japan. waki@gsc.riken.jp
authors.forename: Motoaki
authors.initials: M
authors.lastname: Wakiyama
authors.forename: Reiko
authors.initials: R
authors.lastname: Muramatsu
authors.forename: Yoko
authors.initials: Y
authors.lastname: Kaitsu
authors.forename: Mariko
authors.initials: M
authors.lastname: Ikeda
authors.forename: Shigeyuki
authors.initials: S
authors.lastname: Yokoyama
mesh_terms: D000818:Animals; D002460:Cell Line; D004331:Drosophila melanogaster / Q000166:cytology* / Q000502:physiology*; D007763:Lac Operon / Q000235:genetics*; D056968:Lac Repressors / Q000235:genetics*; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000378:metabolism*
chemical_list: D056968:Lac Repressors; D011994:Recombinant Proteins