Extracting Streptococcus Sanguinis/Porphyromonas Gingivalis Platelet-Aggregating Surface Proteins

To investigate thrombogenic mechanisms of surface proteins on Streptococcus sanguinis(S.s)(PAAP) and Porphyromonas gingivalis(P.g)(Hgp44) through recombinant expression and functional characterization of their platelet-binding functions, with implications for oral microbiota-associated cardiovascular pathologies. The PAAP and Hgp44 genes of S.s 133-79 and P.g ATCC33277 were amplified by PCR, cloned into pUcm-T vector by T-A cloning and subcloned into pET22b expression vector, respectively, and transformed into BL21 (DE3) to construct a recombinant strains, and induced to express the target proteins. In vitro experiments would be performed to analyze the properties of single and combined PAAP/Hgp44 to platelets and their effects on platelet aggregation follow-up. The Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis confirmed that the electrophoretic mobility of PAAP/Hgp44 corresponded to their theoretical molecular weights. Protein sequencing analysis demonstrated high concordance between the obtained peptide mass fingerprints and the theoretical sequences of PAAP/Hgp44. The study pioneers the production of recombinant PAAP in S.s by constructing a haematopoietic expression vector. These findings lay the foundation for elaborating interspecies microbial dynamics in thromboinflammatory cascades, offering mechanistic insights for cardiovascular risk stratification in periodontitis patients.