Abstract:We established a phage display surface protein library of Brucella melitensis 16M strain and to lay a foundation for the screening of a new diagnostic antigen. Recombinant Brucella melitensis 16M strain gene library was constructed by using phagemid vector pYW01. The protein library of B. melitensis 16M strain was constructed infected by helper phage VCSM13. Purified by PEG and enlarged library, the plasmid DNA was randomly extracted from the library to sequence and analyze. The novel proteins are identified by subtractive scanning of the protein library with vaccine serum and infected serum. Complete-enzyme-linked immunosorbent assay (c-ELISA) and Western blot were used to evaluate novel serodiagnostic antigens. B. melitensis 16M strain gene library was constructed by DNA recombination. The library's capacity was 108 pfu / mL with performance randomness. Gene library was infected by helper phage VCSM13 to generate protein library. The plasmid DNA of protein library were randomly extracted to sequence and analyze. The result reveals that phage display protein library of B. melitensis 16M strain was successfully constructed. Six fusion proteins were identified with subscribe scanning and verified by c-ELISA and Western blot.
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