Immunogenicity validation of Burkholderia pseudomallei protein BPSS1622 and its subcellular localization
JIN Haolong
HU Yi
LI Qian
MA Tengfei
HAN Dan
HU Zhiqiang
MAO Xuhu
Abstract：This study was designed to validate immunogenicity of Burkholderia pseudomallei protein BPSS1622 and explore its subcellular localization.The prokaryotic expression vector pET-22b/BPSS1622 was constructed and then expressed in E.coli BL21;the recombinant protein was purified by ion-exchange column chromatography.Then the New Zealand white rabbits were immunized by using the purified recombinant protein for preparation of immune serum.Meanwhile,the immunogenicity of the expressed BPSS1622 protein was confirmed by ELISA and Western blot analysis.Finally,the subcellular localization of BPSS1622 protein was probed by Western blotting.Data showed that BPSS1622 gene was cloned from the genomic DNA of B.pseudomallei and the vector pET-22b/BPSS1622 was constructed successfully;the engineering strain E.coli BL2/BPSS1622 was obtained by transformation of the vector pET-22b/BPSS1622.The molecular weight of expressed protein BPSS1622 was 22 kDa;the titer of the polyclonal antibody was 1 ∶ 1 200 000.Subcellular localization analysis indicated the native protein BPSS1622 localizes in cell wall and membrane of B.pseudomallei;bioinformatics analysis indicated that this protein may has the activity of ATP-dependant RNA helicase.Taken together,the immunogenicity of protein BPSS1622 has primarily confirmed,which lay a foundation for laboratory diagnosis and prevention of melioidosis disease.
Keywords：Burkholderia pseudomalleiBPSS1622 proteinAntiserumSucellular localization
Publication Date：2017-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：6( 771-776 )