Inducible protein expression in Drosophila Schneider 2 cells using the lac operator-repressor system

被引：2

作者：

Wakiyama, Motoaki ^{[1
]}

Muramatsu, Reiko ^{[1
]}

Kaitsu, Yoko ^{[1
]}

Ikeda, Mariko ^{[1
]}

Yokoyama, Shigeyuki ^{[1
,2
]}

机构：

[1] RIKEN Syst & Struct Biol Ctr, Yokohama, Kanagawa 2300045, Japan

[2] Univ Tokyo, Grad Sch Sci, Bunkyo Ku, Tokyo 1130033, Japan

来源：

BIOTECHNOLOGY LETTERS | 2011年 / 33卷 / 12期

关键词：

Drosophila S2 cells; lac operator-repressor; Inducible expression; IPTG; ESCHERICHIA-COLI; TRANSFORMATION; PROMOTER; BINDING; LINES;

D O I：

10.1007/s10529-011-0721-6

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

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.

引用

页码：2361 / 2366

页数：6

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