Purification of p24 protein expressed in Bacillus subtilis and evaluation of its immunogenicity in mice

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Tác giả: Đức Hoàng Nguyễn, Thị Phượng Trang Phan, Thị Tinh Tươm Trương

Ngôn ngữ: vie

Ký hiệu phân loại: 571.961 Physiology and related subjects

Thông tin xuất bản: Tạp chí Phát triển Khoa học & Công nghệ, 2017

Mô tả vật lý:

Bộ sưu tập: Metadata

ID: 492528

Thêm vào giỏ

p24 là một trong những protein cấu thành capsid của Human immunodeficiency virus (HIV). Trong quá trình xâm nhiễm, HIV đưa cả capsid này vào tế bào chủ nhằm thực hiện một số vai trò quan trọng trong các giai đoạn sống của virus trong tế bào chủ. Do đó, protein p24 có tiềm năng ứng dụng trong phát triển vaccine ngăn ngừa HIV. Với định hướng sử dụng p24 làm vaccine, nghiên cứu này tiến hành tạo chủng Bacillus subtilis có khả năng biểu hiện protein p24 và thu nhận protein này. Đây là chủng vi sinh vật an toàn, không gây độc cho người và động vật, đã có hệ thống biểu hiện cho phép biểu hiện vượt mức protein tái tổ hợp lên đến 10-30% so với protein tổng số. Plasmid tái tổ hợp pHT1537 mang gene gagp24 mã hóa cho protein p24 được dung hợp với đuôi LysSN-6His đã được dòng hóa thành công, cho phép biểu hiện protein p24 trong B. subtilis dưới sự cảm ứng của IPTG. Sự hiện diện của protein p24 dung hợp được kiểm tra bằng phương pháp SDS-PAGE. Protein này được thu nhận bằng cột ái lực chứa Ni2+. Đánh giá khả năng tạo kháng thể kháng protein p24 ở chuột bằng thử nghiệm ELISA và Western blot.p24 protein is a component of the HIV particle capsid. It plays an essential role in HIV to infect into the host cell and in the cycle life of virus. Therefore, this protein can be used in the orientative study "to create and produce HIV's vaccine". This study created the new Bacillus subtilis strain which expressed p24 protein. B. subtilis a safety and non-toxic bacteria strain for humans and animals, has system expression to allow over expression recombinant protein up to 10-30 % of total proteins. Plasmid pHT1537 was cloned successfully, containing lysSN-6his-gagp24 gene to encode p24 protein fused with LysSN protein and to allow the expression of p24 protein in B. subtilis by IPTG inducer. The target protein in the cell was cheked by SDS-PAGE. The p24 fused protein was parified from His Trap column which contained Ni2+. Evaluation of the ability to produce antibody against p24 protein in mice by ELISA and Western blot was caried out.

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