鸡AvBD6抗沙门氏菌的表达筛选与抗菌特性-基础兽医学专业论文.docxVIP

AbstractSalmonella pullorum is one of the most serious intestinal bacteria. Screening advantage chicken β-defensins which can resist Salmonella pullorum infection by studying expression regulation in vivo is important.In this study, animal model of chickens infected with Salmonella pullorum was established to screen advantage chicken β-defensins, analyse the structure-activity of the advantage AvBD using bioinformatics software, and improve the biological selection activity by the molecular design. Specific details and results were as follows:Expression screening of resisting against Salmonella pullorum of AvBD6The Roman chickens was inoculated with Salmonella pullorum to established infection model, collected duodenal tissue after infection within 24 h (0 h, 3 h, 6 h, 12 h and 24 h), using real-time quantitative PCR to detect AvBD genes expression patterns during infection ,the results showed that Salmonella pullorum infection within 24 h, AvBD1, 3, and 12 did not detect the change of expression levels significantly; the expression levels of AvBD8, 10, 11, 13 and 14 was significantly (P0.05) decrease at different time; the expression levels of AvBD2, 4, 5, 6, 7and 9 increased significantly at different time periods (P0.05), in which the expression levels of AvBD6 rised with largest rate, and 24 h after infection with the highest expression, so select AvBD6 as the advantage chicken β-defensinof anti-Salmonella pullorum.Structure-activity analysis of antimicrobial quality of AvBD6AvBD6 physicochemical characterization shows that it has the general nature of antimicrobial peptides-cationic and amphiphilic; transmembrane prediction analysis shows AvBD6 can not form a film channel and the target of AvBD6 is likely to the bacterial cytoplasm; secondary structure analysis and spatial model building showed AvBD6 has no α helix and amphiphilic domain, indicating that cell membrane action ability of AvBD6 is relatively poor. Gel retardation assay fully explain AvBD6 can a