Isolation And Identification Of Streptococcus Horse Strangles And FN Binding Protein Immunogenicity Study

In this study, By separating and the identification of Streptococcus horse strangles from Xinjiang.Cloning and prokaryotic expression of Streptococcus horse strangles from Xinjiang FNE and FNEB genes.Getting the FNE and FNEB truncated protein to immunized mice. Analyzing the immunogenicity of theexpressed product.Separating and getting Gram-positive Cocci from the pus in the incidence of horses by the clinicalmanifestations of Streptococcus horse strangles. Which one is Streptococcus equi by biochemicalidentification and PCR identification. It could pathogenic in mice. A fragment of FNE gene and FNEB geneare amplified from Streptococcus equi subsp. equi isolated from Xinjiang by PCR and cloned intopMD19-T vector. After sequence analysis by molecular biology softwares, the truncated FNE gene andFNEB gene are subcloned into the prokaryotic expression vector pET-30a and resulting plasmid istransformed into host BL21(DE3) cells, in which a recombinant protein is expressed by inducing withIPTG．The SDS-PAGE analysis revealed that the expressed recombinant protein are46kD and58kD,Then the expression and antigenicity of the recombinant proteins are detected by Western blot.This study successfully isolated a Streptococcus horse strangles from Xinjiang Yili region. Successfulcloning and expression of Streptococcus horse strangles from Xinjiang FNE and FNEB truncated gene,Obtaining better protective effect of immunity in mice. Lay the foundation for the restructuring horse glandStreptococcus FNE and development FNEB protein subunit vaccine.