The Prokaryotic Expression And Preliminary Application For Recombinant Proteins Of Gene BsSRP And Gene BsABF In Baylisascaris Schroederi

Giant panda is the peculiar valuble endangered animal in China,which is the symbol of World Wildlife Fund.Baylisascaris schroederi parasitize in the gastrointestinal tract of panda specificly and cause the host emaciation,anemia,abdominal pain,vomiting as well as organ inflammation.Baylisascariosis is one of the main lethal factors that cause death of giant panda.However the existing primary diagnosis of Baylisascariosis include brine floatation method and PCR method,which can not be content with the request of celerity,facility with early diagnosis.Not only that,B.schroederi parasitize in the multi-bacterial environment and the promising application of the functional protein has not been explored yet.Until now the promising application of the functional proteins in B.schroederi has not been explored extensively yet.In this study,the gene Serine protease inhibitor(BsSRP)and gene Antibacterial factor(BsABF)was recombinat-expressed,their localization and preliminary application were also studied.Meanwhile the results aid orientations for the early diagnosis and application of functional proteins.1.Prokaryotic expression and primary evaluation of diagnostic value of serine protease inhibitor in Baylisascaris schroederiSerine protease inhibitors superfamily contains numerous members which have different functions and participate in various important physiological activities.Some of them in parasite have been reported as vaccine antigens or diagnostic antigens.In this study BsSRP was recombinant-expressed and as a result,the open reading frame of BsSRP gene(441bp)encoded a 16.06 k Da protein which shared a 61% homology with Toxocara canis;while the result of western-blot revealed that the recombinant protein could be recognized by positive serum of pandas infected with B.schroederi.It mainly distributed in hypodermis,muscle and eggs of parasite;the indirect ELISAs based on BsSRP showed with a sensitivity of 91.67% and specificity of 100%.The recombinant protein has diagnostic value consequently.2.Prokaryotic expression and primary evaluation of antibacterial activity of antibacterial factor in Baylisascaris schroederiAntibacterial peptides are important component in organisms and can play non-specific immune function which are mostly cationic small peptides.Parasites are mainly dependent on the innate immune system as invertebrates.This study amplified BsABF gene from B.schroederi and the open reading frame of BsABF gene(276bp)encoded a 10.01 k Da protein which shared a 94% homology with Ascaris suum;whereafter the western blot results showed that the recombinant protein could be identified by positive serum of pandas infected with B.schroederi.The protein is mainly distributed in hypodermis and intestinal.Meanwhile it is revealed that the recombinant protein has the antibacterial activity.