Prokaryotic Expression of Rabbits Bordetella Bronchiseptica PertactinRecombinant Protein and Development of an Indirect ELISA for Detection of Its Antibodies
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    Abstract:

    Objective To get Bordetella bronchiseptic (Bb)expressing recombinant protein pertactin(PRN)and establish an indirect ELISA for detection of rabbit Bordetella bronchiseptic antibodies. MethodsAccording to a pair of designed specific primers, PRN gene of rabbit Bordetella bronchiseptica was amplified by PCR and inserted into prokaryotic expression vector pGEX-4T-1. The recombinant expression plasmids were transfected into BL21(DE3) strain. The optimal concentration of coated antigen recombinant protein PRN and serum dilution was determined to develop the ELISA technique. ResultsPRN gene of Bordetellabronchiseptica was successfully cloned and expressed. The SDS- PAGE and Western blot analysis unfolded the excellent immunogenicity of the recombinant proteins which were used as coating antigens to develop the ELISA method for Bb-specific antibody diagnosis. The peridium consistency of the recombinant protein PRN determined in this experiment was 500 ng /mL, and the optimal testing serum dilution was 1:40. ConclusionsThe PRN indirect ELISA developed in this study offers a simple and practical way for monitoring antibody of Bb, and provides much information for laying a basis for development of a Bb diagnosis kit. 

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