Nghiên cứu tối ưu các điều kiện biểu hiện tạm thời gen mã hóa kháng nguyên M của virus PRRS trong lá cây thuốc lá Nicotiana benthamiana

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Tác giả: Hoàng Hà Chu, Thị Thương Hồ, Thị Minh Hằng Nguyễn, Thu Giang Nguyễn, Trung Nam Nguyễn, Bích Ngọc Phạm

Ngôn ngữ: vie

Ký hiệu phân loại: 571.962 Physiology and related subjects

Thông tin xuất bản: Công nghệ Sinh học, 2018

Mô tả vật lý: 293-300

Bộ sưu tập: Metadata

ID: 492087

Thêm vào giỏ

Reproductive disorders and respiratory swine (Porcine Reproductive and Respiratory Syndrome - PRRS) is an infectious disease caused by the PRRS virus (PRRSV), spread fast speeds, causing mass death when infected pigs. M protein is one of the major structural protein of the PRRSV, has a molecular weight of about 19 kDa, encoded by the open reading frame 6 (ORF6), which is used to design a subunit vaccine against back PRRSV. In this study, conditions for transient expression of the gene encoded the M protein of PRRSV in leaves of the Nicotiana benthamiana by agroinfiltration method were determined. The results show that optimal conditions for transient expression of protein M in the leaf are used the simultaneous of the A. tumefaciens strain containing vector carrying the gene encoding the protein M and A. tumefaciens containing vector carrying gene encoding supported protein HC-Pro PVY, concentration of acetosyringone with 450 µM, the bacterial cell density used to infect into leaf with OD600 is 0.5, The physiological age of the leaf suitable for infecting bacteria was young leaf and mature leaf of tobacco plants 4 to 6 weeks of age, and the transient expression time of gene encoding protein M of PRRSV in tobacco leaves is most effective 6 days after infecting. This agroinfiltration method can be used to express large amounts of protein - antigen M of PRRSV in tobacco plants to produce vaccines against this virus.Xác định các điều kiện để biểu hiện tạm thời gen mã hóa protein M của virus PRRS trong lá cây thuốc lá Nicotiana benthamiana bằng phương pháp thẩm lọc nhờ Agrobacterium tumefaciens. Kết quả cho thấy, điều kiện tối ưu để biểu hiện protein tạm thời trong lá cây thuốc lá là sử dụng đồng thời chủng A. tumefaciens chứa vector chuyển gen mang gen mã hóa protein M và A. tumefaciens chứa vector hỗ trợ mang gen mã hóa protein HC-Pro PVY
nồng độ chất dẫn dụ và cảm ứng AS 450 μM
mật độ tế bào vi khuẩn được sử dụng để xâm nhiễm vào lá có giá trị OD600 bằng 0,5
tuổi sinh lý của lá phù hợp cho vi khuẩn xâm nhiễm bằng phương pháp hút chân không là lá non và lá bánh tẻ của các cây thuốc lá 4 - 6 tuần tuổi và thời gian biểu hiện tạm thời gen mã hóa protein M của virus PRRS trong lá cây thuốc lá hiệu quả nhất là 6 ngày sau chuyển gen.

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