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TIGHTLY CONTROLLED TETRACYCLINE‐INDUCIBLE TRANSCRIPTION SYSTEM FOR EXPLOSIVE GENE EXPRESSION IN CULTURED SILKWORM CELLS
Author(s) -
Tatsuke Tsuneyuki,
Lee Jae Man,
Kusakabe Takahiro,
Iiyama Kazuhiro,
Sezutsu Hideki,
Uchino Keiro
Publication year - 2013
Publication title -
archives of insect biochemistry and physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.576
H-Index - 66
eISSN - 1520-6327
pISSN - 0739-4462
DOI - 10.1002/arch.21083
Subject(s) - biology , doxycycline , promoter , transgene , tetracycline , transcription (linguistics) , gene , gene expression , microbiology and biotechnology , silencer , regulation of gene expression , drosophila melanogaster , genetics , mechanical engineering , linguistics , philosophy , engineering , inlet , antibiotics
Tetracycline‐inducible gene expression ( T et‐on) system is a particularly powerful tool for transgenic research and has become one of the first choices for the control of transgene expression in a mammal and a fly. Previously, we have generated the modified reverse tetracycline‐controlled transactivators and tet O promoters for a B ombyx mori T et‐on system. In order to further improve this system, G iant, a transcriptional silencer from D rosophila melanogaster, is introduced to repress leaky transcription in the absence of doxycycline. Further, the promoter responsibility to the tetracycline‐controlled transactivators is facilitated by introducing a synthetic minimal core promoter. With the tightly regulated second‐generation silkworm T et‐on system, we obtain up to 300‐fold induction of gene expression with the addition of doxycycline.