布鲁氏菌Cu/Zn SOD蛋白生物信息学分析及原核表达OA

To perform bioinformatics analysis and recombinant expression of Cu/Zn superoxide dismutase(Cu/Zn SOD)protein,a target antigen for Brucellosis vaccines.A range of bioinformatics tools was adopt-ed to predict and analyze the sequence conservation,physicochemical properties,structural features,epitope distribution,and interacting proteins of Cu/Zn SOD.The following tools were employed:NCBI,Protter,Ex-PASy,IEDB,SYFPEITHI,and STRING database,along with SOPMA and AlphaFold software.The ex-pression plasmid pET-28a(+)-sodc was constructed using gene synthesis methods and transformed into BL21(DE3)competent cells.The plasmid was induced for expression using IPTG,and the protein was identified via SDS-PAGE and Western Blot.The protein was purified using an affinity chromatography col-umn.The Cu/Zn SOD protein is comprised of 173 amino acids,with a theoretical molecular weight of 18 130.67 u and an isoelectric point of 6.24.The signal peptide sequence,which spans the first 19 amino acids at the N-terminus,exhibits high conservation among Brucella species in B.abortus,B.melitensis,B.suis,and B.canis.The secondary structure is comprised of 9.83％α-helix,7.51％β-turns,57.23％random coils,and 25.43％extended strand.It harbors multiple T-cell and B-cell antigenic epitopes and has the capa-bility to interact with various proteins.Additionally,the Cu/Zn SOD was successfully constructed,identi-fied,and purified.The Cu/Zn SOD protein of Brucella exhibits high amino acid sequence conservation,sta-bility,and hydrophilicity.It also contains a broad range of B/T cell epitopes,which may enhance its immu-nogenicity,thereby providing a valuable reference for the development of Brucellosis vaccines.