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Sản xuất peptide chống viêm cecropin A trong nội nhũ hạt gạo
Tóm tắt
Cecropin A là một peptide kháng khuẩn tự nhiên có hoạt tính ly giải nhanh chóng, mạnh mẽ và bền vững đối với một loạt các tác nhân gây bệnh, do đó có tiềm năng sinh học lớn trong công nghệ sinh học. Trong nghiên cứu này, chúng tôi báo cáo một hệ thống để sản xuất cecropin A sinh học trong hạt gạo. Các cây gạo biến gen biểu hiện gen cecropin A tổng hợp được tối ưu hóa theo codon, được điều khiển bởi một trình khởi động đặc hiệu cho nội nhũ, cụ thể là trình khởi động glutelin B1 hoặc glutelin B4, đã được tạo ra. Chuỗi peptide tín hiệu từ glutelin B1 hoặc glutelin B4 đã được liên kết với đầu N của trình tự mã hóa cecropin A. Chúng tôi cũng nghiên cứu xem sự hiện diện của tín hiệu giữ lại KDEL trong lưới nội bào có ảnh hưởng đến sự định vị và tích lũy cecropin A trong tế bào hay không. Các cây gạo biến gen cho thấy sự tích hợp gen ổn định và di truyền. Chúng tôi cho thấy cecropin A tích lũy trong các thể protein lưu trữ trong nội nhũ gạo, đặc biệt là trong các thể protein loại II, cho thấy rằng các peptide tín hiệu glutelin ở đầu N đóng một vai trò quan trọng trong việc định hướng cecropin A đến bào quan này, không phụ thuộc vào việc được gán với tín hiệu giữ lại KDEL. Việc sản xuất cecropin A trong hạt gạo biến gen không ảnh hưởng đến khả năng sống của hạt giống hay sự phát triển của cây con. Hơn nữa, hạt giống cecropin A biến gen cho thấy khả năng chống lại sự nhiễm trùng của các tác nhân gây bệnh nấm và vi khuẩn (Fusarium verticillioides và Dickeya dadantii, tương ứng), cho thấy rằng cecropin A được sản xuất trong cây có hoạt tính sinh học. Hạt gạo có khả năng duy trì sản xuất cecropin A sinh học và tích lũy trong các thể protein. Hệ thống này có thể mang lại lợi ích cho việc sản xuất chất chống vi khuẩn này cho các ứng dụng tiếp theo trong bảo vệ cây trồng và bảo quản thực phẩm.
Từ khóa
#cecropin A #peptide kháng khuẩn #gạo biến gen #nội nhũ #tín hiệu KDELTài liệu tham khảo
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