凡纳滨对虾WAP基因重组表达及抗病功能分析.

[Objective] Because of the serious disease problem of Iitopenaeus vannamei, it is important to study the disease resistance genes of L. vannamei. In this study, the WAP gene of L. vannamei was cloned and its tissue expression and anti-disease function were analyzed to provide reference for the development of anti-disease drugs for L. vannamei. [Method] The specific primers of W AP gene were designed, and the expression and distribution of the W AP gene in 11 tissues including blood cells, eye stem, gills, hepatopancreas, heart, stomach, muscle, nerve, intestine, cecum and epidermis were detected by RT-PCR. The W AP expression in gills and intestinal tissues of L. vannamei was detected by fluorescence quantitative PCR. The recombinant W AP protein was produced and the binding ability of recombinant W AP to LPS, PGN and LTA was analyzed by ELISA. [Result] It was found that WAP gene was expressed in all 11 tissues tested, with the highest expression in blood cell and the lowest expression in epidermis, eye stalk and hepatopancreas. The W AP gene expression was significantly increased in gill and intestinal tissues of infected L. vannamei compared with that of normal L. vannamei ( P < 0. OS) . The expression of WAP gene in the gill and intestinal tissues of infected L. vannamei were as 5. 9258 and 1. 5147 times as that of healthy L. vannamei,respectively. The recombinant W AP fusion protein could bind to bacterial polysaccharides LPS, PGN and LTA directly, and the order of binding ability was PGN > LPS > LTA. [Conclusion] The WAP gene was closely related to the disease resistance and immune function of L. vannamei, W AP fusion protein has potential in developing anti white feces drugs for L. vannamei. [ABSTRACT FROM AUTHOR]