Nghiên cứu sử dụng promoter Pgrac57 để tạo vectơ tự biểu hiện mang chỉ thị GFP cho Bacillus subtilis

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Tác giả: Tran Thi Anh Dao, Thi Hanh Au, Thanh Dung Dang, Duc Hoang Nguyen, Thuy My Linh Nguyen, Thi Phuong Trang Phan, Thi Lan Truong

Ngôn ngữ: vie

Ký hiệu phân loại: 570.78 Life sciences Biology

Thông tin xuất bản: Tạp chí Khoa học (Trường Đại học Sư phạm TP. Hồ Chí Minh), 2018

Mô tả vật lý: 100-108

Bộ sưu tập: Metadata

ID: 492254

Thêm vào giỏ

Bacillus subtilis có nhiều ưu điểm như: An toàn, không gây bệnh, không có nội độc tố nên được sử dụng nhiều trong sản xuất protein trị liệu và enzyme công nghiệp quan trọng. Tiềm năng ứng dụng B. subtilis cho biểu hiện protein tái tổ hợp được nâng cao trong những năm gần đây với sự ra đời của vectơ cảm ứng pHT sử dụng promoter Pgrac. Trong nghiên cứu này, chúng tôi phát triển hệ thống vectơ biểu hiện không cần chất cảm ứng cho B. subtilis dựa trên promoter Pgrac57. Kết quả đã xây dựng được vectơ tự biểu hiện pHT1686 điều hòa bởi promoter Pgrac57, cho phép biểu hiện GFP mà không cần chất cảm ứng trong chủng B. subtilis 1012, lượng biểu hiện protein GFP đạt mức 11% protein tổng số của tế bào và hoạt tính protein GFP ngang bằng với hệ thống có cảm ứng.Bacillus subtilis has many advantages such as: safe, non-pathogenic, endotoxinfree features. Therefore, B. subtilis has been widely ultilized in therapeutic proteins and important industrial enzymes production. Over the last few years, the potential application of B. subtilis for recombinant proteins synthesis has been significantly enhanced by using pHT - vector system with Pgrac promoter. In this study, we developed inducer-free expression vector for B. subitlis based on Pgrac57 promoter and used GFP protein as a reporter. The results showed that we successfully constructed inducer-free vector pHT1686 with Pgrac57 promoters. These vectors are totally able to express GFP protein without any inducer in B. subtilis strain 1012. Moreover, GFP expression level reached 11% of the total cellular proteins. Additionally, GFP activities, are in the same range with the inducer-free vector.

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