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Ngày xuất bản: 13-06-2025
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XÂY DỰNG VÀ PHÁT TRIỂN BIOSENSOR TỪ TẾ BÀO NẤM MEN ỨNG DỤNG TRONG PHÁT HIỆN CÁC HỢP CHẤT GÂY ĐỘT BIẾN GEN
,
Nguyễn Hữu Đức
1, 2, 3
,
Nguyễn Đức Thắng
1, 2, 3
,
Nguyễn Công Hiếu
1, 2, 3
Từ khóa
Biosensor, độc tính gen, độc tính tế bào, gen thông báo (reporter gene), GFP (green fluorescent protein), methyl methansulfonate, menadione
Tóm tắt
Biosensor được phát triển nhờ việc thiết kế và xây dựng một plasmid thông báo mang gen GFP được điều khiển bởi promoter RNR2. Plasmid thông báo được biến nạp vào tế bào nấm men, sau đó được xử lý với các nồng độ MMS và menadione khác nhau. Thông qua cơ chế cảm ứng hoạt động bởi hợp chất gây biến đổi gen MMS, mà promoter RNR2 điều khiển sự phiên mã và biểu hiện của GFP, dẫn đến tín hiệu huỳnh quang phát ra khi đo tại 485nm (sóng kích thích) và 535nm (sóng phát xạ). Cường độ huỳnh quang đo được tỷ lệ thuận với nồng độ MMS có trong môi trường phân tích và đạt cực đại tại 1mM, nhưng giảm dần khi xử lý với nồng độ MMS trên 1mM, hiệu ứng này là do hoạt động hô hấp tế bào và quá trình sinh tổng hợp ATP bị kìm hãm. Ngược lại, menadione không gây kích thích promoter RNR2 hoạt động, nên không có tín hiệu huỳnh quang được ghi nhận. Chính vì vậy, biosensor theo thiết kế này không những có tiềm năng trong việc xác định, phát hiện các hợp chất gây biến đổi gen, độc hại tới con người và môi trường, mà còn phát hiện các hợp chất mới có cùng tính chất hóa học với MMS và có thể ứng dụng trong phát hiện thuốc mới và sàng lọc dược phẩm.
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