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| Establishment of a tetracycline-off and heat shock-on gene expression system in tobacco |
| ZHOU You, LI Jin-hua, PAN Yu, ZHENG Yu, PAN Yang-lu, DING Yu-mei, SU Cheng-gang, ZHANG Xing-guo |
| Key Laboratory of Horticulture Science for Southern Mountainous Regions, Ministry of Education/College of Horticulture and Landscape Architecture, Southwest University, Chongqing 400715, P.R.China |
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Abstract The tetracycline (Tet)-off gene expression regulation system based on the TetR-VP16/Top10 construct has not been widely utilized in plants, for its highly expressed TetR-VP16 activator is toxic to some plants and repeatedly replenishing tetracycline to turn off the constitutively active system is a tedious process. To solve these problems, a Tet-off and heat shock (HS)-on gene expression regulation system was constructed in this study. This system is composed of a chimeric transactivator gene TetR-HSF that is derived from a Tet repressor (TetR) and a HS transcription factor (HSF) controlled by a HS promoter HSP70m, and a Tet operator containing hybrid promoter, Om35S, that drives expression of the β-glucuronidase (GUS) gene. The resultant system yields a GUS expression pattern similar to that of the HSP70m promoter under inducing temperatures and at 35 and 40°C drives GUS expression to a similar level as the Cauliflower mosaic virus (CaMV) 35S promoter. Further examination revealed that the TetR-HSF and GUS genes were induced by HS, reaching peak expression after 1 and 6 h treatment, respectively, and the HS induction of the expression system could be inhibited by Tet. This system will provide a useful tool for transgenic studies of plants in the laboratory and in the field, including transgene function analysis, agronomic trait improvement, biopharmaceutical protein production and others.
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Received: 11 July 2016
Accepted:
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| Fund: The study was supported by the National High-Tech R&D Program of China (2010AA10060705) and the Transgenic Engineering Crops Breeding Special Funds from China’s Ministry of Agriculture (2009ZX08010-005B). |
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Corresponding Authors:
ZHANG Xing-guo, Tel: +86-23-68250974, Fax: +86-23-68251274, E-mail: zhangxg63@163.com
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Cite this article:
ZHOU You, LI Jin-hua, PAN Yu, ZHENG Yu, PAN Yang-lu, DING Yu-mei, SU Cheng-gang, ZHANG Xing-guo.
2017.
Establishment of a tetracycline-off and heat shock-on gene expression system in tobacco. Journal of Integrative Agriculture, 16(05): 1112-1119.
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