The Study Of Sequence Analysis And Adhesion Activity Of Actinobacillus Pleuropneumoniae

Actinobacillus pleuropneumoniae (Actinobacillus pleuropneumoniae, APP) pleuropneumoniae is caused by disease pathogens, widely popular in the world, and is a serious infectious disease in the pig industry against important infectious diseases.2009, Auger, etc. APP lung epithelial cells in vitro adhesion experiments, we found a novel gene up-regulated expression, and Haemophilus influenzae Hsf adhesin from a transfer of high homology; in many bacteria in the adhesin from a transfer have some protective immunity, which has developed from a transfer out of adhesin and efficient commercialization of the vaccine, the most typical is the pertussis bacilli pertactin (PRN) as a vaccine component to add, pertactin is the vaccine increased antibody levels to reduce important factor in pertussis infection. Because APP has a trimer in the same adhesin from a transfer, so try to use case studies of pertussis vaccine. APP has a large number of serotypes, their virulence and immunogenicity of different, mainly among non-immune cross-serotype protection, restricting the pathogenic mechanism of APP and bottlenecks in vaccine research, a serious impact on APP's diagnosis, quarantine and prevention. Search for effective diagnostic, quarantine and prevention methods, this study chose APP serum 2,4,6,8,9,10 type strain study, APP reported to GenBank since the transfer of serum type 5b adhesin gene (CP000569.1500018-510061bp) primers by PCR method to find the gene, bioinformatic analysis and prediction of the serotype trimeric adhesin from a transfer function areas, to find a complete serum 8 functional areas and functional areas of its name adh gene cloning and sequence analysis of the adh gene expression, while the mice immune protection against APP infection for determination, verification adh protective recombinant fusion protein, and with thrombin to remove the GST fusion protein tags, Adh protein purification, and to detect whether the Adh protein adhesion activity of lung epithelial cells. The results showed that serum 2,4,6,8,9,10-type adhesin from a transfer of genes with large differences, found that 8 in serum and serum type 5b adhesin amino acid sequence from a transfer of 92% homology, amino acids deduced sequences of 82%; and successful fusion protein GST-8Adh, experiments show that the APP protein and antibody can inhibit the adhesion of the role of lung epithelial cells, and protective power of serum 4,8-type of 9/10, protection against other serotypes is not obvious, but there are also some protection. This study demonstrated in APP trimeric adhesin from a transfer can be used as genetic engineering vaccine candidate targets, and different serotypes of APP for the future of new multivalent vaccine research, the establishment of pathogenic mechanism to provide the necessary foundation.