Preparation And Characterization Of Campylobacter Jejuni Peb1-mediated Intestinal Rbcg Vaccine With High Affinity

BCG (Bacillus Calmette Guerin) was found to be able to induce Th1 predominant response in long-term clinical application and experimental study [1]. More than this, many of the BCG ingredients are capable of inducing Th1 predominant response alone. Hence we suppose that if we transfect allergen Der p2 gene into BCG, the body will present the allergen as a BCG ingredient after it is immunized, consequently required immune status might be induced by appropriate vaccination. Our research group's former studies have shown that Der p2 can be expressed in prokaryotic expression system, and its amount of protein could reach 20% of total [2,3]. In order to improve the affinity of rBCG with intestinal mucosa, we chose Campylobacter jejuni outer membrane protein PEB1 as a medium by cloning it into Der p2-rBCG using recombinant BCG technology, and identified it for further enhanced vaccine efficacy. ObjectiveTo construct the recombinant BCG, on which was able to express house dust mite antigen Der p2 and C.jejuni PEB1 in form of lipoprotein for oral asthma vaccine, the pCW-Der p2-PEB1-rBCG..Methods and ResultsThe pCW-Der p2 and pUC-PEB1 plasmid DNA were amplified and recombinant plasmid pCW-Der p2-PEB1 was synthesized. The recombinant plasmid was transferred into competent BCG, after that rBCG positive bacterium strains were selected by hygromycin resistance screening. Target gene was detected in positive rBCG by specific amplification using PCR. rBCG was checked by indirect immunofluorescence with anti-Der p2 monoclonal antibody and antiserum against mouse PEB1.ConclusionRecombinant BCG pCW-Der p2-PEB1-rBCG Expressing Der p2-PEB1 Fusion Protein on its Surface was successfully constructed and this product may provide basis for the further research of genetic engineering vaccine.