tailieunhanh - PHÁT HIỆN PROTEIN P74 TRÊN VI-RÚT GÂY BỆNH ĐỐM TRẮNG (WHITE SPOT SYNDROME VIRUS) Ở TÔM: PROTEIN TIỀM NĂNG TẠO VẮC-XIN

Vi-rút gây bệnh đốm trắng (WSSV) là một trong những vi-rút gây bệnh nguy hiểm trên tôm và là mối đe dọa lớn cho nghề nuôi trồng thủy sản. Tuy nhiên, chưa có biện pháp hữu hiệu để điều trị bệnh đốm trắng. Hiện nay nhiều nghiên cứu chỉ ra rằng protein VP28 và VP19 là những protein giúp tạo vắc-xin protein đề kháng lại sự xâm nhiễm của WSSV cho tôm. Phương thức lan truyền bệnh qua đường miệng (peroral infectivity) luôn luôn cần sự hiện diện của các nhân tố truyền bệnh qua đường miệng (peroral infectivity factor – PIFs). . | Tạp chí Khoa học 2012 22c 119-128 Trường Đại học Cần Thơ PHÁT HIỆN PROTEIN P74 TRÊN VI-RÚT GÂY BỆNH ĐỐM TRẮNG WHITE SPOT SYNDROME VIRUS Ở TÔM PROTEIN TIỀM NĂNG TẠO VẮC-XIN Trần Thị Mỹ Duyên1 Peng Ke2 và Just M. Vlak2 ABSTRACT White spot syndrome virus WSSV is one of the most devastating viral diseases in shrimp and a major threat in the shrimp aquaculture industry. Currently there are no comprehensive strategies demonstrate that white spot disease can be treated. Many scientific research efforts have shown that a protein vaccine including the envelope proteins VP28 and VP19 as a target for intervention can be a possible treatment to protect WSSV infection. Peroral infectivity factors PIFs are absolutely required for oral infectivity not only in Baculoviruses but most likely also in other large invertebrate circular double stranded ADN viruses. These PIFs could be alternative targets for immune-intervention. Recent computational investigations indicate that WSSV also has so-called per os infectivity factors. In this report a recombinant bacterium was generated by inserting a fragment of the gene WSSV ORF 72 encoding for a putative WSSV P74 protein in expression plasmid pET28a. Expression of a recombinant WSSV P74 protein fragment of 66 kDa was performed in E. coli. Protein expression level was optimized by various approaches to obtain the highest level for high yield protein purification. Polyclonal antibodies were raised in rabbit against the isolated WSSV P74 protein. These antibodies specifically reacted against a 108 kDa protein and some other smaller proteins which is the size expected from a full-length P74 protein. During antibody generation Penaeus vannamei P. vannamei were infected with WSSV for virus isolation. After that antibodies will be used to detect recombinant bacterial WSSV P74 protein and used to further investigate the location of P74 on the surface ofWSSV virions and for virus neutralization experiments. This result shows that P74 protein was .