tailieunhanh - Tinh sạch và đánh giá hoạt tính sinh học của kháng nguyên P54 tái tổ hợp của virus gây bệnh tả lợn Châu Phi (African swine fever virus) biểu hiện trên cây thuốc lá Nicotiana benthamina

Trong nghiên cứu này, trình tự ectodomain p54 của chủng VNUAHY-ASFV/Vietnam2019 được tối ưu mã biểu hiện trên Nicotiana benthamiana, sau đó được tổng hợp nhân tạo và chèn vào vector pRTRA có chứa các motif trimer, oligomer. | TNU Journal of Science and Technology 228 13 314 - 321 PURIFICATION AND EVALUATION OF THE BIOACTIVITY OF THE RECOMBINANT P54 ANTIGEN OF THE AFRICAN SWINE FEVER VIRUS EXPRESSED FROM Nicotiana benthamina Nguyen Thi Tra1 2 Nguyen Thu Giang1 Ho Thi Thuong1 2 Trinh Thai Vy1 2 Hoang Thi Thu Hang1 2 Vu Duy Thai Son1 Le Thi Tra My1 Nguyen Thi Thu Hien1 Le Thi Thanh1 Pham Bich Ngoc1 2 1Institute of Biotechnology VAST 2Graduate University of Science and Technology - VAST ARTICLE INFO ABSTRACT Received 24 7 2023 African swine fever is a dangerous infectious disease caused by ASFV African swine fever virus in pigs with mortality rates up to 100 . Currently no officially Revised 30 8 2023 licensed commercial vaccine against African swine fever is on the market. p54 is Published 13 9 2023 considered one of the main target proteins for ASFV vaccine development and ASFV diagnostic kit production. In this study the DNA sequence encoding the ectodomain of p54 of the VNUAHY-ASFV Vietnam2019 strain was optimized for KEYWORDS expression in Nicotiana benthamiana then artificially synthesized and inserted into Transient expression the pRTRA vector containing the trimer and oligomer motifs. Next the entire expression cassettes were inserted into the pCB301 vector and successfully A. tumefaciens transformed into Agrobacterium tumefaciens for transient expression in Nicotiana N. benthamiana benthamiana. The expression of recombinant p54 protein in plants was determined ASFV strain in Vietnam by Western blot. The better expression motif was selected for purification by Immobilized affinity chromatography tested for the immune response in mice and Recombinant p54 protein evaluated for ASFV-specific IgG antibody titer by ELISA reaction. P54-pII-tp was selected to purify and induce an immune response in mice. ELISA results showed that recombinant p54-pII-tp stimulated ASFV-specific antibody production in laboratory mice. This result is a premise for research and development of subunit vaccines