| Products/Services Used | Details | Operation |
|---|---|---|
| Plasmid DNA Preparation | A plasmid construct containing a mutated, synthetic S. pseudintermedius spEX (designed as described below) with BamHI/NotI cloning sites, was obtained commercially (Genscript Piscataway, NJ USA) (Table 1).The native SpEX open reading frame (ORF) (705bp) excluding N-terminal signal peptide was amplified from S. pseudintermedius strain 06–3228 genomic DNA and the mutant SpEX was amplified from a pUC19-spEX-M plasmid (Genscript Piscataway, NJ USA) (Table 1) using taq polymerase (rTaq, Takara). The endotoxin concentration in purified recombinant SpEX-M was measured using a ToxinSensor Chromogenic LAL Endotoxin Assay Kit (Genscript). | Get A Quote |
| PCR and Cloning | Get A Quote |
Staphylococci have evolved numerous strategies to evade their hosts' immune systems. Some staphylococcal toxins target essential components of host innate immunity, one of the two main branches of the immune system. Analysis of the Staphylococcus pseudintermedius secretome using liquid chromatography mass spectrometry guided by genomic data, was used to identify an S. pseudintermedius exotoxin provisionally named SpEX. This exoprotein has low overall amino acid identity with the Staphylococcus aureus group of proteins named staphylococcal superantigen like proteins (SSLs) and staphylococcal enterotoxin- like toxin X (SEIX), but predictive modeling showed that it shares similar folds and domain architecture to t... More
