Objective:　To　construct　a　recombinant　fusion　protein　with　pneumococcal　surface　protein　A　(PspA)　of　Stretococcus　pneumonia　(SPIN)　family　[clade　1　and　2,　and　to　analyze　the　immunogenici-Iv　of　the　fusion　protein.　Methods:　The　gene　fragments　encoding　the　α-helix　of　PspA　of　the　two　clades　were　amplified　by　PCR　and　then　inserted　into　the　expression　vector　pET-27b(+)　to　construct　the　recombinant　expression　plasrnid.　The　transformed　Escherichia　coli　BL21　strains　carrying　expression　plasmid　were　induced　by　IPTC　to　express　the　recombinant　protein.　The　titers　and　affinity　of　antibodies　against　PspA　protein　were　measured　by　EIJSA.　An　opsonophagocytic　assay　and　an　animal　experiment　were　performed　to　evaluate　the　immunogenicity　of　the　recombinant　protein.　Results:　Double　enzyme　cutting　and　gene　sequencing　confirmed　the　two　purpose　gene　fragments　were　correctly　expressed　in　the　expression　vector　pET-27b(+).　The　titers　of　anti-PspA　antibody　in　the　serum　of　Kunming　(KM)　mice　immunized　with　the　fusion　protein　were　1　×　104.　The　affinity　of　anti-PspA　antibody　reached　to　2　×　105.　The　rates　of　recombinant　PspA6B-PspA05　protein　mediated　phagocytosis　for　SPN6B,　SPN05　and　SPN01　strains　were　20%,　15%　and　8.8%,　respectively.　No　SPIN23K　strain　was　engulfed　by　macrophages　upon　the　stimulation　with　PspA6B-PspA05　protein.　The　survival　tales　of　mice　injected　with　SPN05,　SPN6B,　SPN01　and　SPN23F　strains　were　respectively　75%,　92%,　75%　and　33%　upon　the　immunization　of　PspA6B-PspA05　protein.　Conclusion:　The　recombinant　fusion　protein　PspA6B-PspA05,　constructed　with　the　PspA　proteins　of　Stretococcus　pneumonia　family　I　clade　1　and　2,　was　successfully　expressed　in　the　E.　coli　prokaryotic　system　with　the　advantage　of　high　immunogenicity.　High　titers　of　anti-PspA　antibodies　with　high　specificity　were　induced　in　KM　mice　upon　the　stimulation　with　Ps-pA6B-PspA05　protein.　Moreover,　a　cross-protective　immunity　was　induced　in　KM　mice　upon　the　immunization　with　PspA6B-PspA05　protein.　Copyright　©　2015　by　the　Chinese　Medical　Association.