Establishment of a tetracycline-off and heat shock-on gene expression system in tobacco
Autor: | Yanglu Pan, You Zhou, Yu-mei Ding, Yu Zheng, Yu Pan, Xingguo Zhang, Jin-hua Li, Cheng-gang Su |
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Rok vydání: | 2017 |
Předmět: |
0106 biological sciences
0301 basic medicine Agriculture (General) Transgene Plant Science TetR-HSF transactivator tobacco 01 natural sciences Biochemistry S1-972 03 medical and health sciences chemistry.chemical_compound Transactivation Food Animals Gene expression TetR Gene Regulation of gene expression Ecology biology Activator (genetics) fungi food and beverages biochemical phenomena metabolism and nutrition biology.organism_classification Molecular biology 030104 developmental biology chemistry tetracycline-off heat shock-on Animal Science and Zoology Cauliflower mosaic virus gene expression system Agronomy and Crop Science 010606 plant biology & botany Food Science |
Zdroj: | Journal of Integrative Agriculture, Vol 16, Iss 5, Pp 1112-1119 (2017) |
ISSN: | 2095-3119 |
Popis: | 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. |
Databáze: | OpenAIRE |
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