Tạo dòng, biểu hiện và tinh sạch Protein A-L2 bằng silica

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Tác giả: Thanh Tấn Nguyễn, Thị Thùy Trinh Nguyễn, Văn Hiếu Trần

Ngôn ngữ: Vie

Ký hiệu phân loại: 576 Genetics and evolution

Thông tin xuất bản: Khoa học (Đại học Cần Thơ) 2022

Mô tả vật lý: 45517

Bộ sưu tập: Metadata

ID: 403401

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Tóm tắt
Chủ đề

Các hạt silica (SiO2) được cố định protein là vật liệu đầy triển vọng trong các lĩnh vực y sinh hay cảm biến sinh học. Những nghiên cứu trước cho thấy protein ribosome L2 của vi khuẩn Escherichia coli liên kết mạnh với các hạt silica. Ngoài ra, protein A là thụ thể bề mặt có tính ổn định cao, có nguồn gốc từ vi khuẩn Staphylococcus aureus, sự liên kết giữa protein A và kháng thể được xem là một trong những tương tác protein-protein kinh điển được nghiên cứu nhiều nhất hiện nay. Nghiên cứu này được tiến hành nhằm tạo tiểu phần protein A dung hợp với protein L2 tái tổ hợp bằng cách cấu trúc vector pET22b-proAx1-L2. Protein được biểu hiện thông qua hệ thống E. coli BL21(DE3) và được kiểm tra bằng SDS-PAGE. Dựa vào sự liên kết đặc hiệu với hạt silica, Protein A-L2 được tinh sạch bằng các hạt silica trần không biến tính và sử dụng dung dịch MgCl2 nồng độ cao để dung ly protein mục tiêu. Phương pháp tinh sạch bằng hạt silica sẽ mang lại hiệu quả như tinh sạch nhanh chóng, tiết kiệm chi phí và độ tinh sạch tương đối., Tóm tắt tiếng anh, Protein immobilized silica (SiO2) particles are promising materials in the fields of biomedicine or biosensors. Through previous studies, it was found that the Escherichia coli ribosomal protein L2 strongly binds to silica particles. In addition, protein A which is derived from Staphylococcus aureus bacteria, is a highly stable surface receptor. The interaction between protein A and antibodies is considered one of the classical protein-protein interactions. This study created a fusion protein A subunit with recombinant L2 protein by constructing the vector pET22b-proAx1-L2. The Protein A-L2 was expressed in E. coli BL21(DE3) system and analyzed using SDS-PAGE. Based on the specific binding to silica particles, Protein A-L2 was purified by unmodified bare silica particles and a high concentration of MgCl2 solution. With the purification method by silica beads, high efficiencies were achieved, such as rapid purification, cost savings, and comparative purity.

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