Using the Tet-on system to construct n inducible transposon in plant
Date Issued
2008
Date
2008
Author(s)
Weng, Ruei-Chien
Abstract
In this study, the Tet-on system from microorganism was intergrated with Ac transposon in an attempt to construct an efficient and inducible transposon system in plant. We combined the Ac transposase with the inducible Tetpro promoter based on Tet-on system to create the inducible transposon, TetAc. As a control, the inducible Tetpro promoter was also fused with luc gene, termed as TetLUC system. Each system was introduced into rice and tobacco by agro-transformation. The transgenic calli of TetAc system were induced with several concentrations of doxycycline and induction periods. These inductions were applied to the control, TetLUC system. We found about 88.8 percent spontaneous transposition of TetAc in the transgenic rice lines. The Tetpro promoter of the TetLUC was also observed for about 90.47 percent spontaneous expression in transgenic rice lines. However, the Tetpro promoter of the TetLUC system could be induced by doxycycline and increased for about 2 ~ 92.7 -fold and 2 ~ 24.43 -fold of luc gene expression ratio (compare with uninduced transgenic line) in the rice and tobacco calli, respectively. In order to establish a double control system “TetPR”, we used luc and rtTA gene to fuse with the Tetpro and PR-1a inducible promoter, respectively. The TetPR system was indroduced into rice and tobacco by agro-transformation. No spontaneous expression was detected. After induced with doxycycline and salicylic acid in the rice calli of TetPR system, the LUC expression of the rice calli could achieve 16.82 -fold compared with non-induction. However, the luc gene expression is relatively low compared with the luc gene which was driven by 35S promoter. Further experiments should find a way which can increase the expression of luc gene and the availability of the inducible transposon system in the future.
Subjects
inducible transposon
Tet-on system
transposition
Type
thesis
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