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Bulletin of Botanical Research ›› 2016, Vol. 36 ›› Issue (6): 917-924.doi: 10.7525/j.issn.1673-5102.2016.06.016

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Establishment of A Chemical-inducible Gene Expression System in Tobacco

DAI Li-Juan1, ZHENG Tang-Chun1, LIU Cai-Xia1, LIU Yi1, YOU Xiang-Ling2, QU Guan-Zheng1   

  1. 1. State Key Laboratory of Tree Genetics and Breeding, Northeast Forestry University, Harbin 150040;
    2. College of Life Science, Northeast Forest University, Harbin 150040
  • Received:2016-06-06 Online:2016-11-15 Published:2016-11-16
  • Supported by:
    Program for New Century Excellent Talents in University(No.NCET-12-0808);The National Natural Science Foundation of China(No.31370661)

Abstract: Chemical-inducible expression system is a powerful tool for plant functional genomics and plant genetic engineering applications. XVE is a chemical-inducible expression system using estrogen for chemical induction. The detailed report about XVE using tobacco as plant material was not available yet. In this study, GFP was amplified from the plasmid pROKⅡ-GFP that was preserved in this experiment by PCR. Estrogen inducible plant expression vector(pER8-GFP) was constructed and transformed into wild-type tobacco by Agrobacterium-mediated leaf disc transformation. The positive transformants were screened with hygromycin followed by a further PCR identification. The transformants were induced with different concentrations and different times of estrogen. The expression of GFP in transgenic tobacco was tested by using qRT-PCR and NightSHADE plants in vivo imaging system, and the results showed that the best suitable concentration of estrogen which induced the expression of pER8-GFP in tobacco was 25 μmol·L-1, and the best suitable period was 48 h. At the same time, the GFP gene appeared in the hypocotyl and root tip cells of transgenic tobacco. It showed that the XVE chemical induction system in tobacco could also be used to control the expression of the target gene efficiently by the induction of estrogen. Our study will contribute to the XVE study of tobacco in future.

Key words: estradiol-inducible, gene expression, XVE, Nicotiana tabacum

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