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  • Công bố khoa học và công nghệ Việt Nam

62.37

Công nghệ sinh học công nghiệp

Bùi Văn Ngọc, Nguyễn Thị Thu Huyền, Mai Chí Thành, Hoàng Thị Yến(1), Nguyễn Thị Hoài Thu, Phạm Thùy Linh, Nghiêm Ngọc Minh, Nguyễn Hữu Đức

Tạo vector biểu hiện mang gen chỉ thị GFP điều khiển bởi promoter RAD54 để tạo Biosensor phát hiện hợp chất gây đột biến gen

Construction of expression vector incorporating a GFP reporter gene under the control of RAD54 promoter for development of biosensor used for detection of genotoxic chemical compounds

Công nghệ Sinh học

2014

3

431-438

1811-4989

Biosensors have been known as potentially powerful analytical tools for determination of the genotoxic potential of chemical compounds which are harmful to human and environment. In the present study, biosensor for detection of genotoxins was developed by design and construction of a reporter plasmid containing GFP gen under regulation of RAD54 promoter. This reporter plasmid was transformed in yeast cells which were then treated with different concentrations of 4-NQO and menadione. Due to induction of genotoxic compounds, 4-NQO at concentration of 0,5 uM triggers the activation of RAD54 promoter that regulates transcription and expresion of downstream gene, GFP, leading to fluorescence signal measured at 485 nm (excitation wave length) and 535 nm (emission wave length). Fluorescence instensity was directly proportional to 4-NQO concentrations when yeast cells treated with less than 1 uM in the environment, but decreasing when exposed to higher 4-NQO concentrations. In contrast, promoter RAD54 was not induced by menadione resulting in no fluorescence signal. Thus, such biosensor is able to identify new and uncharacterized compounds which possess the same chemical properties as 4-NQO and could be used for further development of later biosensors for new drug discovery or in drug screening.

TTKHCNQG, CVv 262