Cloning, expression, purification and antigenic evaluation of hyaluronidase antigenic fragments recombinant protein of streptococcus pyogenes
Streptococcus pyogenes produce an extracellular hyaluronidase which is associated with the spread of the organism during infection. Enzyme hyaluronidase is capable of degrading hyaluronic acid. The aim of the present study was to clone and express antigenic regions of the hylA of S.pyogenes in Escherichia coli. The antigenic regions of hylA gene was inserted in pGEX-4T-1 vector. E. coli BL21 was transformed with hylA_pGEX-4T-1 and gene expression was induced by isopropyl-beta- Dthiogalactopyranoside (IPTG). The expressed protein was purified by affinity chromatography with GST sepharose resin. The integrity of the product was confirmed by western-blot analysis using sera of infected individual. Sera reactivity of infected individuals was further analyzed against the recombinant hyaluronidase protein. Our data shows production of recombinant hyaluronidase improved by pGEX- 4T-1 in Ecoli.
Key word: Hyaluronidase gene, cloning, expression of recombinant gene, antigenic region.